AU2017384681B2

AU2017384681B2 - Monoclonal anti-alpha-synuclein antibodies for preventing tau aggregation

Info

Publication number: AU2017384681B2
Application number: AU2017384681A
Authority: AU
Inventors: Karina FOG; Jeppe Falsig Pedersen
Original assignee: H Lundbeck AS
Current assignee: H Lundbeck AS
Priority date: 2016-12-22
Filing date: 2017-12-21
Publication date: 2024-06-13
Anticipated expiration: 2037-12-21
Also published as: RU2019118922A3; KR102584663B1; US20180179273A1; JP7300990B2; JP2020504729A; CO2019006059A2; CN110114088B; BR112018017172A2; WO2018115225A1; RU2760334C2; KR20190098976A; CL2019001735A1; IL267559A; CA3046857A1; AU2017384681A1; EP3558364A1; US10364286B2; CN110114088A; MX2019007480A; RU2019118922A

Abstract

The invention relates to a novel use of monoclonal anti-alpha-synuclein antibodies. The antibodies can be used to prevent tau aggregation and thereby treating tauopathies such as Alzheimer's disease.

Description

Monoclonal Anti-Alpha-Synuclein Antibodies for Prevent ing Tau Aggregation

Field of invention 5 [0001] The present invention relates to a novel use of monoclonal antibody that immunospecifically bind to alpha-synuclein, as well as to methods of using these molecules and their alpha-synuclein binding fragments in the treatment of tau-pathies such as Alzheimers disease (AD).

10 Reference To Sequence Listing

[0002] This application includes one or more Sequence Listings pursuant to 37 C.F.R. 1.821 etseq., which are disclosed in both paper and computer-reada ble media, and which paper and computer-readable disclosures are herein in corporated by reference in their entireties. 15 Background of the invention

[0003] Age-related neurodegenerative diseases such as Alzheimer's disease (AD) and dementia are one of the largest societal challenges today. The World Health Organization estimates that costs for care of the elderly will con 20 tinue to increase and that the number of diagnosed dementia cases will triple by 2050 (World Health Organization and Alzheimer's Disease International Status Report (2012) DEMENTIA: A public health priority, WHO). The first treatments for AD were neurotransmitter modulators such as acetylcholine esterase inhibitors and NMDA modulators. These therapies became available 25 at the turn of the millennium and still form the cornerstone for symptomatic re lief of memory deficits related to dementia and AD. However, these drugs do not target the underlying causes of AD, accumulation of amyloid-p (AP) pep tide and tau protein aggregates and associated loss of neuronal synapses and eventually neurons. 30 [0004] Longitudinal, community-wide studies of the elderly (Weiner, M.W. et al. (2014) ADNI; Breteler, M.M. et al. (1992) Neuroepidemiology 11 Suppl 1, 23-28; Launer, L.J. (1992) Neuroepidemiology 11 Suppl 1, 2-13) together with large genome-wide association studies (Lambert, J.C. et al. (2013) Nat. Genet. 45, 1452-1458) have shown that AD is a heterogeneous mix of de mentias where up to 10 percent of the advanced AD patients lack amyloid pa thology (Crary, J.F. et al. (2014) Acta Neuropathol. 128, 755-766). Further 5 more, seminal pathological studies by Braak & Braak (Braak, H. and Braak, E. (1996) Acta Neurol. Scand. Suppl 165, 3-12) demonstrated a clear correla tion between the degree of neurofibrillary tangle pathology and cognitive state prior to autopsy. These observations have been reinforced by several investi gators (Nelson, P.T. et al. (2012) J. Neuropathol. Exp. Neurol. 71, 362-381), 10 and in recent longitudinal biomarker studies, which indicate that cerebrospinal fluid (CSF) levels of tau and phospho-tau increase throughout early and late stages of the disease (Jack, C.R., Jr. et al. (2013) Lancet Neurol. 12, 207 216).

[0005] As indicated above, the microtubule-associated protein, tau, and its 15 hyper-phosphorylated version, form the main constituent of intracellular neu rofibrillary tangles, which are one of the main hallmarks of AD. Furthermore, specific genetic variants of tau are associated with familial forms of fronto temporal dementia (FTD). Appearance of tau pathology in AD occurs in a dis tinct spatial pattern, starting in the entorhinal cortex, followed by hippocampal 20 and cortical areas (Braak, H. and Braak, E. (1996) Acta Neurol. Scand. Suppl 165, 3-12). The specific stage of tau pathology also correlates well with cogni tive abilities (Nelson, P.T. et al. (2012) J. Neuropathol. Exp. Neurol. 71, 362 381; Braak, E. et al. (1999) Eur. Arch. Psychiatry Clin. Neurosci. 249 Suppl 3, 14-22). Taken together, this evidence forms the basis of a tau-based hypoth 25 esis for AD. It entails that the intracellular accumulation of tau leads to micro tubule degeneration and spinal collapse. As a result, communication between neurons malfunctions and cell death follows. Recently, it has also been shown that tau itself may form an endo-pathogenic species that can transmit neurodegeneration from one cell to the next (Clavaguera, F. et al. (2009) Nat. 30 Cell Biol. 11, 909-913).

Tau As An Endo-Pathogen

[0006] Clavaguera and colleagues have demonstrated that tau itself may act as an endo-pathogen (Clavaguera, F. et al. (2009) Nat. Cell Biol. 11, 909 5 913). Low spin brain extracts were isolated from P301S tau transgenic mice (Allen, B. et al. (2002) J. Neurosci. 22, 9340-9351), diluted and injected into the hippocampus and cortical areas of young ALZ17 mice. The ALZ17 mouse is a tau transgenic mouse line which only develops late pathology (Probst, A. et al. (2000) Acta Neuropathol. 99, 469-481). The injected ALZ17 mice 10 quickly developed solid filamentous pathology, and administration of immuno depleted brain extracts from P301S mice or extracts from wild type mice did not induce tau pathology. Fractionation of the brain extracts in soluble (Si) and sarcosyl-insoluble tau (P3) (Sahara, N. et al. (2013) J. Alzheimer's. Dis. 33, 249-263) and injection of these into ALZ17 mice demonstrated that the P3 15 fraction is most competent in inducing pathology. It contains most of the intra cellular hyper-phosphorylated filamentous tau. The majority of pathology could also be induced when injecting P301S extracts into the brains of wild type mice, but no NFTs were formed. In subsequent studies, Clavaguera et al. have shown that human tau extracted from post-mortem brain tissue of 20 other tauopathies (Argyrophilic Grain Disease (AGD), Progressive Supranu clear Palsy (PSP), and Corticobasal Degeneration (CBD)) may also induce tau pathology in the ALZ17 model (Clavaguera, F. et al. (2013) Proc. NatI. Acad. Sci. U.S.A. 110, 9535-9540). Since the presentation of these data, sev eral other tau seeding and spreading models have been reported (Ahmed, Z. 25 et al. (2014) Acta Neuropathol. 127, 667-683; Walker, L.C. et al. (2013) JAMA Neurol. 70, 304-310). The main conclusion from these studies indicates a mechanism by which pathogenic tau in intracellular inclusions is secreted from the cell into the periplasmic space. The pathological tau material is then transported along the vesicular sheath in both anterograde and retrograde di 30 rection and subsequently taken up by neighboring cells by means of bulk en docytosis. This mechanism explains why the spread of pathology observed in human disease follows a distinct anatomical pattern. Intriguingly, peripheral administration of pathological tau may accelerate the formation of tau pathol ogy in ALZ17 mice (Clavaguera, F. et al. (2014) Acta Neuropathol. 127, 299 301).

5 Relation between Alpha-synuclein and Tau patholgy

[0007] Alpha-synuclein is a member of a family of proteins including beta- and gamma-synuclein and synoretin. Alpha-synuclein is expressed in the normal state associated with synapses and is believed to play a role in regulating synaptic vesicle release and thereby affect neural plasticity, learning and 10 memory.

[0008] Several studies have implicated alpha-synuclein with a central role in Parkinson's Disease (PD) pathogenesis. The protein can aggregate to form intracellular insoluble fibrils in pathological conditions. For example, synuclein accumulates in Lewy Bodies (LB) (Spillantini et al., Nature (1997) 388:839 15 40; Takeda et al., J. Pathol. (1998) 152:367-72; Wakabayashi et al., Neuro sci. Lett. (1997) 239:45-8). Mutations in the alpha-synuclein gene as well as duplications and triplications of the gene co-segregate with rare familial forms of parkinsonism (Kruger et al., Nature Gen. (1998) 18:106-8; Polymeropou los, et al., Science (1997) 276:2045-7).

20 [0009] An important finding has been that alpha-synuclein can be secreted into and be present in plasma and cerebrospinal fluid (CSF). Several studies, for example by Pacheco et al. (2015) and others (Conway et al., (2000) Proc Natl Acad Sci USA, 97:571-576; Volles et al., J. Biochem. (2003) 42:7871-7878) have suggested that extracellular-synuclein plays a pathogenic role in the 25 brain. They demonstrated that alpha-synuclein possesses neurotoxicity toward brain neuronal plasma membranes exposed directly to extracellular-synuclein oligomers. Another intriguing hypothesis based on the data of synuclein secre tion is that a prion-like spread of alpha-synuclein underlies the progression of Parkinson's disease and other synucleinopathies (Lee et al., Hansen et al. 30 (2011) J. Clin Invest 121:715-725). These finding have given rise to a hope that extracellular-synuclein could be targeted by immunotherapy (Vekrellis et al. (2011) Lancet Neurol 10:1015-1025) and be a potential treatment of alpha synucleinopathies. In addition to mutations, alternative splicing of the alpha synuclein gene and posttranslational modifications of the protein, such as phosphorylation, ubiquitination, nitration, and truncation can create alpha synuclein protein forms that have enhanced capacity to form aggregated and/or 5 toxic forms of alpha-synuclein (Beyer and Ariza, Mol Neurobiol. 2013 Apr;47(2):509-24). However, the precise pathological species of alpha-synu clein in alpha-synucleinopathies remains unknown. Various misfolded/aggre gated/secreted species ranging from oligomers to fibrils, and different post translational modifications have been associated with toxicity but there is no 10 consensus on which is most important, if indeed there even is a single toxic species.

[00010] The co-appearance of pathologies, for example Lewy bodies, Abeta plaques and neurofibrillary tangles in subsets of patients with PD or Lewy bodies in a subset of AD patients (Galpern and Lang, Ann. Neurol. (2008), 15 59: 449-458) has led to investigations of to what extent aggregation prone proteins can cross-seed each other. Alpha-synuclein and tau proteins have been reported to be able to induce fibrillization of each upon co-incubation in vitro (Giasson et al., (2003) Science 300: 636-640). In cellular systems there is both evidence supporting and not supporting a cross-seeding of tau with al 20 pha-synuclein fibrils. Holmes et al., could not demonstrate a cross-seeding of tau with fibrils made from full-length alpha-synculein in a FRET based tau re porter cell line (Holmes et al., (2014) PNAS doi/10.1073: E4376-E4385). Nor could Tau aggregation be induced with fibrillated full-length alpha-synuclein A53T or a PTA-precipitated (to enrich for fibrillated alpha-synuclein) brain 25 sample from a multiple system atrophy (MSA) patient (Woerman et al., (2015) PNAS doi/10.1073: E4949-E4958. Others have reported that under some conditions alpha-synuclein can induce tau aggregation, for example alpha synuclein fibrils made from N-terminal truncated (21-140) alpha-synuclein was shown to induce tau phosphorylation in QB1293 cells (Waxman and 30 Giasson (2011) J. Neurosci 31: 7604-7618). In neuronal cultures full length fibrillated alpha-synuclein do seed tau aggregation. However fibrillated alpha synuclein made from truncated alpha-synuclein (1-120 or 32-140) can through repeated self-seeding in cells (5 or 10% of fibrillated alpha-synuclein from each passage included as seeds in the fibrilization of the subsequent passage) (Guo el at., (2013) Cell 154: 109-117).

[00011] To our knowledge no studies has hypothesised that secreted oligo meric or fibrillated forms of alpha-synuclein could be a contributing factor in 5 the early pathogenesis of AD or other tauophaties in general independent of visible alpha-synuclein inclusions. The reports that could demonstrate a cross-seeding of tau with alpha-synuclein have focussed on explaining why there in some PD patients, with manifest alpha-synculein aggregation (Lewy bodies) are found neurofibrillary tangles. These studies speculate that the 10 cross-seeding can take place in areas where there is a close physiological association of tau and alpha-syncuelin deposits (Giasson et al., (2003) Sci ence 300: 636-640; Waxman and Giasson (2011) J. Neurosci 31: 7604-7618; Guo el at., (2013) Cell 154: 109-117). The idea that soluble extracellular oli gomeric/fibrillated forms of alpha-synuclein and not intracellular aggregates in 15 the form of Lewy bodies or Lewy neurites are the important contributing factor in the generation of tau neurofibrillar tangles is new.

Alpha-synuclein immunotherapies

[00012] Antibodies binding to alpha-synuclein have been developed as potential 20 therapeutic agents to treat synucleinopathies, also known as Lewy body dis eases (LBDs). Synucleinopathies are characterized by deposition of intracel lular protein aggregates microscopically visible as Lewy bodies (LBs) and/or Lewy neurites, where the protein alpha-synuclein is a major component (Jellinger, Mov Disord. 2012 Jan;27(1):8-30; McKeith et al., Neurology (1996) 25 47:1113-24). Synucleinopathies include Parkinson's disease (including idio pathic Parkinson's disease) and Diffuse Lewy Body (DLB) disease (also known as Dementia with Lewy Bodies (DLB), Lewy body variant of Alzhei mer's disease (LBV), Combined Alzheimer's and Parkinson disease (PD), pure autonomic failure and multiple system atrophy (MSA; e.g., Olivoponto 30 cerebellar Atrophy, Striatonigral Degeneration and Shy-Drager Syndrome).

[00013] Several different antibodies to alpha-synuclein have been shown to have therapeutic effect in preclinical animal models. Both an antibody targeting an epitope involving alpha-synuclein residues 91-99 and antibodies targeting an epitope that involves alpha-synuclein residues 118-126 have been shown to have an effect on motor and cognitive deficits in transgenic mice (Games et al. 2014). The most advanced of these antibodies is a humanized antibody based on the mouse monoclonal antibody 9E4, which targets an epitope that 5 involves alpha-synuclein residues 118-126, and which is now in clinical trials in phase 1. Also an antibody that targets an amino-terminal epitope of alpha synuclein has been shown to have possible therapeutic potential in prevent ing spreading and toxicity of pathology in a mouse prion like transfer model (Tran et al. 2014) and a C-terminal antibody 274 which targets an epitope that 10 involves alpha-synuclein residues 120-140 (Bae et al. 2012) was also shown to have an effect in preclinical model on spreading of the pathology from cell to cell. In addition to these, antibodies targeting conformational species such as oligomers and fibrils of alpha-synuclein have been shown to be able to at least reduce the levels of these presumably toxic alpha-synuclein species 15 (Lindtsrbm et al. 2014 and Spencer et al. 2014). These conformational anti bodies that lower alpha-synuclein oligomer levels in vivo, such as mab47 were also shown to target epitopes in the C-terminus of alpha-synuclein, from amino acid 121-125 (US20120308572). Other conformational, fibril and oligo mer specific antibodies also target C-terminal sequences (Vaikath et al. Neu 20 robiol Dis. 2015 Apr 30;79:81-99). Importantly none of these alpha-synuclein antibodies has been claimed to be able to prevent tau aggregation and as a consequence be able to potentially treat tauopathies.

[00014] In this invention we surprisingly discovered that aggregated/fibrillated alpha-synuclein can induce aggregation of Tau and that several antibodies 25 generated to bind alpha-synuclein are able to prevent this aggregation. We show that a panel of different alpha-synuclein antibodies are all able to pre vent aggregation of Tau in the cellular model: An antibody (GM37) that can bind to the presumed toxic alpha-synuclein fragment 1-119/122 (binding to amino acids 112-117 of alpha-synuclein) and neutralize this truncated form of 30 alpha-synuclein, an antibody (2E6) that bind to amino acid 136-140 of alpha synuclein, an antibody (GM63) that bind to amino acid 126-138 of alpha synuclein and an antibody 9E4 that bind to amino acid 118-126 of alpha synuclein. To support that fibrillary forms of alpha-synuclein may contribute to early AD pathology we demonstrate the presence of fibrillated alpha-synu clein in brains from AD patients independent of the presence of Lewy body pathology. This supports that soluble extracellular forms of fibrillated alpha synuclein can potentially play a role in contributing to Tau-pathology in tauopathies such as all AD patients and not only in those characterised with visible alpha-synuclein aggregates (determined by brain imaging or post mor tem staining).

Summary of the invention

[00015] In a first aspect, the present invention relates to the use of an al pha-synuclein binding monoclonal antibody, or an antigen-binding fragment thereof, in the manufacture of a medicament for inhibiting aggregation of tau, wherein said alpha-synuclein binding antibody or antigen-binding fragment thereof is selected from the group consisting of (A)-(D):

(A) a monoclonal antibody, or antigen-binding fragment thereof, comprising: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:2; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6;

(B) a monoclonal antibody, or antigen-binding fragment thereof, comprising: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:33; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6;

(C) a monoclonal antibody, or antigen-binding fragment thereof, comprising: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:34; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6;

(D) a monoclonal antibody, or antigen-binding fragment thereof, comprising:

8a

(a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:35; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6.

[00015a] In a second aspect, the present invention relates to a method of inhibiting aggregation of tau which comprises administering to a subject an effec tive amount of an alpha-synuclein binding monoclonal antibody, or an antigen binding fragment thereof, wherein said alpha-synuclein binding antibody or anti gen-binding fragment thereof is selected from the group consisting of (A)-(D):

8b

(D) a monoclonal antibody, or antigen-binding fragment thereof, comprising:

8c

[00015b] In a third aspect, the present invention relates to a pharmaceutical composition comprising the monoclonal antibody, or an antigen-binding frag ment thereof, as recited in any one of the first or second aspects, and a phar maceutically acceptable carrier.

[00015c] The present invention relates to the use of an alpha-synuclein bind ing monoclonal antibody for inhibiting aggregation of tau.

[00016] The antibodies of the invention, as disclosed herein and in the claims, can be used in treating patients with Alzheimer's disease or patients with a taupahty such as Argyrophilic Grain Disease (AGD), Psychosis, particularly Psychosis due to AD or Psychosis in patients with AD, psychiatric symptoms of patients with Lewy body dementia, Progressive Supranuclear Palsy (PSP), Frontotem poral dementia (FTD or variants thereof), TBI (traumatic brain injury, acute or chronic), Corticobasal Degeneration (CBD), Picks Disease, Primary age-re lated tauopathy (PART), Neurofibrillary tangle-predominant senile dementia, Dementia pugilistica, Chronic traumatic encephalopathy, stroke, stroke recov ery, neurodegeneration in relation to Parkinson's disease, Parkinsonism linked to chromosome, Lytico-Bodig disease (Parkinson-dementia complex of Guam), Ganglioglioma and gangliocytoma, Meningioangiomatosis, Postencephalitic parkinsonism, Subacute sclerosing panencephalitis, Huntington's disease,

8d lead encephalopathy, tuberous sclerosis, Hallervorden-Spatz disease and lipofuscinosis. More typically, the taupathy is selected from the group consist ing of Alzheimer's disease, Argyrophilic Grain Disease (AGD), Psychosis, par ticularly Psychosis due to AD or Psychosis in patients with AD, psychiatric symptoms of patients with Lewy body dementia, Progressive Supranuclear

8e

Palsy (PSP), Frontotemporal dementia (FTD or variants thereof), TBI (trau matic brain injury, acute or chronic), Corticobasal Degeneration (CBD), and Picks Disease.

5 Brief description of figures

[00017] Fig. 1 shows immunization protocols for generation of hybridomas. The table outlines the differences of the immunogens and mouse strains used for the identification of GM37 and GM285. Different HCo17-Balb/c and HCo12/Balb/c mice were immunized independently (description of these mice 10 are provided below). The hybridoma expressing GM37 was identified from mice immunized with full length alpha-synuclein containing amino acids 1-140 fibrils and boosted with truncated alpha-synuclein fragments 1-60 and 1-119 of full length (FL) alpha-synuclein (SEQ ID NO 10). The hybridoma expressing antibody GM285 came from an immunization protocol in which HCo12-Balb/c 15 mice were immunized with full length monomeric alpha-synuclein, amino acids 1-140 followed by a boost with full length fibrillary alpha-synuclein (Example 1).

[00018] Fig. 2 (PANEL A-C) shows screening of GM37 for binding to alpha synu clein, alpha-synuclein homologs and orthologs.

[00019] A) Binding of antibody GM37 to alpha-synuclein using a no wash solu 20 tion based ELISA (FMAT).

[00020] B) Using SPR (Fortebio) binding of antibody GM37 is specific for al pha-synuclein (Alpha Panel) and does not bind the other related synuclein fam ily proteins, beta-synuclein (Beta Panel) and gamma-synuclein (Gamma Panel). Measurements were performed using SPR (Fortebio Octetred) GM37 25 shows similar binding to alpha-synuclein from cynomolgus monkey (Cyno Panel) and mouse (Mouse Panel). (Example 1).

[00021] C) Using SPR (Fortebio Octetred) binding of antibody GM285 is specific for alpha-synuclein and does not bind the other related synuclein family pro teins, beta-synuclein and gamma-synuclein. Measurements were performed using SPR (Fortebio Octetred) shows similar binding of GM285 to alpha-synu clein from cynomolgus monkey (Cyno) and mouse (Mouse)(Example 1).

[00022] Fig. 3 (Panels A-C) shows real time binding Affinity of GM37

[00023] A) Binding of antibody GM37 to alpha-synuclein measured in RU (Rel 5 ative Units) (y-axis) over time (X-axis) as determined by SPR (BlAcore@3000). Goat anti-human IgG was immobilized on the CM5 chip. GM37 was captured on the Goat anti-human IgG immobilized chip and series of concentrations of human alpha-synuclein (3.125, 6.25, 12.5, 25, 50, 100 nM) were tested on binding to the surface. The sensor surface was regenerated between each cy 10 cle.

[00024] B) Signal from binding at different concentrations converted into a binding curve.

[00025] C) Calculated binding constants of antibody GM37 (denoted hgG1 6004-037-C106S) (Example 2).

15 [00026] Fig. 4 (Panels A-C) shows real time binding Affinity of GM285

[00027] A) Binding of antibody GM285 to alpha-synuclein measured in RU (y axis) over time (X-axis) as determined by SPR (BAcore@ 3000). Goat anti human IgG was immobilized on the CM5 chip. GM285 was captured on the Goat anti-human IgG immobilized chip and series of concentrations of human 20 alpha-synuclein (3.125, 6.25, 12.5, 25, 50, 100 nM) were tested on binding to the surface. The sensor surface was regenerated between each cycle.

[00028] B) Signal from binding at different concentrations converted into a binding curve.

[00029] C) Calculated binding constants of antibody GM285 (denoted hIgG1 25 6004-285) (Example 2).

[00030] Fig. 5 (Panels A-C) shows real time binding of comparator antibody 9E4

[00031] A) Shows binding of 9E4 to alpha-synuclein measured in RU (y-axis) over time (X-axis) as determined by SPR (BAcore@ 3000). Goat anti-human IgG was immobilized on the CM5 chip. 9E4 was captured on the chip by its binding to Goat anti-human IgG that had been immobilized to the chip. A series 5 of concentrations of human alpha-synuclein (3.125, 6.25, 12.5, 25, 50, 100 nM) were tested for binding to the surface. The sensor surface was regenerated between each cycle.

[00032] B) Signal from binding at different concentrations converted into a binding curve.

10 [00033] C) Calculated binding constants for antibody 9E4. (Example 2).

[00034] Fig. 6 (Panels A-B) shows epitope mapping of antibody GM37 and GM285. ELISA data showing relative levels of binding of the antibodies to se quential peptides (20mers) derived from alpha-synuclein amino acid sequence 95 - 132 (the other nonbinding peptides are not shown).

15 [00035] A) GM37 epitope requires peptide sequence ILEDMP (SEQ ID NO:9) for full binding.

[00036] B) GM285 requires peptide ILED (SEQ ID NO:19) for full binding. (Ex ample 3).

20 [00037] Fig 7 shows a table comparing the binding rate kinetic parameters of GM37 and variants 1-3 to immobilized recombinant human alpha-synuclein. The binding was measured using SPR and the rates were determined using a 1:1 binding algorithm (BlAcore@ T200).

25 [00038] Fig. 8 Tau aggregation induced by alpha-synuclein seeds prevented by alpha-synuclein antibodies. Figure 7 upper panel shows that tau aggregation can be efficiently induced by alpha-synuclein seeds (fibrillated recombinant alpha-synuclein) in a type of cellular model commonly used to assess the ef fect of agents interfering with Tau aggregation. The alpha-synuclein seed in duced tau aggregation can be prevented by alpha-synuclein antibodies in general - exemplified by 9E4 (Elan, Prothena) and Lundbeck antibodies HLD1, GM37 ("37") and GM63 ("63"). Antibodies against Tau phosphorylated 5 on Serine 396 (D1.2, C10.2 and humanized (h) c10.2 and another tau anti body denoted Lu0041G) and control antibodies with no affinity for alpha-synu clein have no effect on tau aggregation, supporting the importance of the therapeutic antibody to interact with the seeding species, not the endogenous protein (Example 12). Figure 8 lower panel shows an outline of the aggrega 10 tion assay in HEK293 cells. Cells are transfected with cDNA encoding human full length Tau with the P301L mutation. Twentyfour hour's later cells are treated with alpha-synuclein seeds in combination with antibodies. After 48 hours the level of Tau aggregation is measured in cell homogenates using a biochemical assay (Example 5). 15

[00039] Fig 9 Presence of alpha-synuclein aggregates in frontal cortex from all 50 AD cases - group divided in mid-stage (Braak ll/IV) and late-stage (Braak V/VI) AD. 2 DLB samples are included as control (two upper lines in A). No detection of alpha-synuclein-serinel29 phosphorylation in AD samples. De 20 mentia with Lewy body (DLB) samples are included positive as controls. A and B demonstrate the presence of aggregated alpha-synuclein in frontal cor tex of 50 AD patients measured by a biochemical method. Patients where clinically diagnosed with AD and the diagnosis confirmed by postmortem his tological staining for Tau and Abeta. None of the patients presented with 25 Lewy body pathology (aggregated serinel29 phosphorylated alpha-synu clein), figure 9C. The presence of alpha-synuclein aggregates in all patients and the absence of serine29 phosphorylated alpha-synuclein (a marker for Lewy bodies), supports the hypothesis that aggregated forms of alpha-synu clein is present in all AD patients before alpha-synuclein pathology might 30 manifest as Lewy bodies. We suggest that these aggregated - pre-Lewy body forms of alpha-synuclein can act as a contributing factor in inducing tau pathology (figure 8). In summary we hypothesize that any alpha-synuclein an tibody that are capable of neutralizing alpha-synuclein aggregates (seeds) or by other means prevent alpha-synuclein aggregates in entering neurons or glia cells and facilitate aggregation of Tau, will have a therapeutic potential to treat tauopathies.

5 [00040] DETAILED DESCRIPTION OF THE INVENTION Definitions

[00041] As used herein, the term "alpha-synuclein" is synonymous with "the al pha-synuclein protein" and refers to any of the alpha-synuclein protein isoforms (identified in, for example, UniProt as P37840, 1-3). The amino acid 10 numbering of alpha-synuclein is given with respect to SEQ ID NO:10 as shown below, with methionine (M) being amino acid residue: SEQ ID NO:10: MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYV GSKTKEGVVHGVATVAEKTKEQVTNVGGAVVTGVTAVAQK 15 TVEGAGSIAA ATGFVKKDQL GKNEEGAPQE GILEDMPVDP DNEAYEMPSEEGYQDYEPEA

[00042] The present invention relates to antibodies and to fragments of antibod ies that are capable of immunospecifically binding to alpha-synuclein, and in particular to human alpha-synuclein, and in one embodiment exhibit the abil 20 ity to immunospecifically bind to an epitope within amino acids 110-140 of hu man alpha-synuclein. According to some embodiments, the antibodies bind to an epitope within amino acids 112-117, 112-115, 118-126, 126-138 or 136 140 of human alpha-synuclein.

[00043] By the term "taupathy" is typically referred to as neurodegenerative dis 25 eases associated with the pathological aggregation of tau. Typically, the tau pathy is selected from the group consisting of Alzheimer's disease, Argyrophilic Grain Disease (AGD), Psychosis, particularly Psychosis due to AD or Psycho sis in patients with AD, psychiatric symptoms of patients with Lewy body de mentia, Progressive Supranuclear Palsy (PSP), Frontotemporal dementia 30 (FTD or variants thereof), TBI (traumatic brain injury, acute or chronic), Corti cobasal Degeneration (CBD), Picks Disease, Primary age-related tauopathy

(PART), Neurofibrillary tangle-predominant senile dementia, Dementia pugilis tica, Chronic traumatic encephalopathy, stroke, stroke recovery, neurodegen eration in relation to Parkinson's disease, Parkinsonism linked to chromosome, Lytico-Bodig disease (Parkinson-dementia complex of Guam), Ganglioglioma 5 and gangliocytoma, Meningioangiomatosis, Postencephalitic parkinsonism, Subacute sclerosing panencephalitis, Huntington's disease, lead encephalo pathy, tuberous sclerosis, Hallervorden-Spatz disease and lipofuscinosis. More typically, the taupathy is selected from the group consisting of Alzheimer's dis ease, Argyrophilic Grain Disease (AGD), Psychosis, particularly Psychosis due 10 to AD or Psychosis in patients with AD, psychiatric symptoms of patients with Lewy body dementia, Progressive Supranuclear Palsy (PSP), Frontotemporal dementia (FTD or variants thereof), TBI (traumatic brain injury, acute or chronic), Corticobasal Degeneration (CBD), and Picks Disease. In particular, the tauopathies may be selected from Alzheimer's disease, Argyrophilic Grain 15 Disease (AGD), Psychosis due to AD or Psychosis in patients with AD, and psychiatric symptoms of patients with Lewy body dementia.

[00044] The term "antibody" (Ab) in the context of the present invention refers to an immunoglobulin molecule or according to some embodiments of the inven tion a fragment of an immunoglobulin molecule which has the ability to specif 20 ically bind to an epitope of a molecule ("antigen"). Naturally occurring anti bodies typically comprise a tetramer which is usually composed of at least two heavy (H) chains and at least two light (L) chains. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region, usually comprised of three domains (CH1, CH2 25 and CH3). Heavy chains can be of any isotype, including IgG (IgG1, IgG2, IgG3 and IgG4). Each light chain is comprised of a light chain variable region (abbreviated herein as VL) and a light chain constant region (CL). Light chains include kappa chains and lambda chains. The heavy and light chain variable region is typically responsible for antigen recognition, while the heavy 30 and light chain constant region may mediate the binding of the immunoglobu lin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system. The VH and VL regions can be further subdivided into regions of hy pervariability, termed "complementarity determining regions," that are inter spersed with regions of more conserved sequence, termed "framework re gions" (FR). Each VH and VL is composed of three CDR Domains and four 5 FR Domains arranged from amino-terminus to carboxy-terminus in the follow ing order: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an an tigen. Of particular relevance are antibodies and their epitope-binding frag ments that have been "isolated" so as to exist in a physical milieu distinct 10 from that in which it may occur in nature or that have been modified so as to differ from a naturally occurring antibody in amino acid sequence.

[00045] As used herein, the term "epitope-binding fragment of an antibody" means a fragment of an antibody capable of immunospecifically binding to an epitope. An epitope-binding fragment may contain 1, 2, 3, 4, 5 or all 6 of the 15 CDR Domains of such antibody and, although capable of immunospecifically binding to such epitope, may exhibit an immunospecificity, affinity or selectiv ity toward such epitope that differs from that of such antibody. Preferably, however, an epitope-binding fragment will contain all 6 of the CDR Domains of such antibody. An epitope-binding fragment of an antibody may be a sin 20 gle polypeptide chain (e.g., an scFv), or may comprise two or more polypep tide chains, each having an amino-terminus and a carboxyl terminus (e.g., a diabody, an Fab fragment, an Fab 2 fragment, etc.). Fragments of antibodies that exhibit epitope-binding ability can be obtained, for example, by protease cleavage of intact antibodies. More preferably, although the two domains of 25 the Fv fragment, VL and VH, are encoded by separate genes, such gene se quences or their encoding cDNA can be joined, using recombinant methods, by a flexible linker that enables them to be made as a single protein chain in which the VL and VH regions associate to form monovalent epitope-binding molecules (known as single-chain Fv (scFv); see e.g., Bird et al. , (1988) Sci 30 ence 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. (U.S.A.) 85:5879-5883). Alternatively, by employing a flexible linker that is too short (e.g., less than about 9 residues) to enable the VL and VH regions of a single polypeptide chain to associate together, one can form a bispecific antibody, diabody, or similar molecule (in which two such polypeptide chains associate together to form a bivalent epitope-binding molecule) (see for instance PNAS USA 90(14), 6444-8 (1993) for a description of diabodies).Examples of the epitope-binding fragments encompassed within the present invention include 5 (i) a Fab'or Fab fragment, a monovalent fragment consisting of the VL, VN, CL and CH1 domains, or a monovalent antibody as described in W02007059782; (ii) F(ab')2 fragments, bivalent fragments comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd frag ment consisting essentially of the VH and CH1 domains; (iv) a Fv fragment 10 consisting essentially of a VL and VH domains, (v) a dAb fragment (Ward et al., Nature 341, 544-546 (1989)), which consists essentially of a VH domain and also called domain antibodies (Holt et al; Trends Biotechnol. 2003 Nov;2i(II) :484-90); (vi) camelid or nanobodies (Revets et al; Expert Opin Biol Ther. 2005 Jan;5_(I) : 111-24) and (vii) an isolated complementarity determin 15 ing region (CDR). Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they may be joined, using re combinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain antibodies or single chain Fv (scFv), see 20 for instance Bird et al., Science 242, 423-426 (1988) and Huston et al., PNAS USA 85, 5879-5883 (1988)). These and other useful antibody fragments in the context of the present invention are discussed further herein. It also should be understood that the term antibody, unless specified otherwise, also includes antibody-like polypeptides, such as chimeric antibodies and human 25 ized antibodies, and antibody fragments retaining the ability to specifically bind to the antigen (antigen-binding fragments) provided by any known tech nique, such as enzymatic cleavage, peptide synthesis, and recombinant tech niques. An antibody as generated can possess any isotype. As used herein, "isotype" refers to the immunoglobulin class (for instance IgG1, IgG2, IgG3, 30 IgG4) that is encoded by heavy chain constant region genes. Such antibody fragments are obtained using conventional techniques known to those of skill in the art; suitable fragments capable of binding to a desired epitope may be readily screened for utility in the same manner as an intact antibody.

[00046] The antibody GM37, 37 or GM37wt (used interchangeable herein) is in tended to mean an antibody or antigen-binding fragment thereof comprising or consisting of the Heavy Chain as given in CDR1-3 SEQ ID Nos 1-3 and the Light Chain CDR1-3 as given in SEQ ID Nos 4-6. 5 [00047] The variants 1, 2 and 3 of GM37 differs from GM37 by difference(s) in the CDR 2 sequence in the heavy chain as given in: GM37 Variant 1 Heavy chain CDR2 SEQ ID NO: 33 GM37 Variant 2 Heavy chain CDR2 SEQ ID NO: 34 GM37 Variant 3 Heavy chain CDR2 SEQ ID NO: 35 10 [00048] The antibody GM285 or IgG-6004-285(used interchangeable herein) is intended to mean an antibody or antigen-binding fragment thereof comprising or consisting of the Heavy Chain as given in CDR1-3 SEQ ID Nos 20-22 and the Light Chain CDR1-3 as given in SEQ ID Nos 23-25.

[00049] The antibody GM63 or 63 (used interchangeable herein) is intended to 15 mean an antibody or antigen-binding fragment thereof comprising or consist ing of the Heavy Chain as given in CDR1-3 SEQ ID Nos 51-53 and the Light Chain CDR1-3 as given in SEQ ID Nos 54-56.

[00050] The antibody 9E4 is intended to mean an antibody or antigen-binding fragment thereof comprising or consisting of the Heavy Chain as given in 20 CDR1-3 SEQ ID Nos 44-46 and the Light Chain CDR1-3 as given in SEQ ID Nos 47-49.

[00051] The antibody 2E6 or m2E6 is intended to mean an antibody or antigen binding fragment thereof comprising or consisting of the Heavy Chain as given in CDR1-3 SEQ ID Nos 62-64 and the Light Chain CDR1-3 as given in 25 SEQ ID Nos 65-67.

[00052] The variants of 2E6, ch2E6, 2E6-HLD1, 2 or 3, has differences in their Heavy Chain and Light Chain outside the CDR regions as compared to 2E6

[00053] ch2E6 comprises or consist of a Heavy Chain SEQ ID NO: 70 and comprises or consist of a Light Chain SEQ ID NO: 71. 30 [00054] 2E6-HLD-1 comprises or consist of a Heavy Chain SEQ ID NO: 72 and comprises or consist of a Light Chain SEQ ID NO: 73.

[00055] 2E6-HLD-2 comprises or consist of a Heavy Chain SEQ ID NO: 74 and comprises or consist of a Light Chain SEQ ID NO: 75.

[00056] 2E6-HLD-2 comprises or consist of a Heavy Chain SEQ ID NO: 76 and comprises or consist of a Light Chain SEQ ID NO: 77.

[00057] The affinity matured forms of HLD1: 7A10, 5A1, 9D7, 9G11, 7C4, L3, 5 8D9, 9C12 or 6B6 has differences in their CDR regions as defined in the se quence listing and claims compared to 2E6.

[00058] The antibody "'6B6" is intended to mean an antibody consisting of the Light Chain SEQ ID NO 120 and Heavy Chain SEQ ID NO 121.

[00059] The antibody "5A1" is intended to mean an antibody consisting of the Light 10 Chain SEQ ID NO 104 and Heavy Chain SEQ ID NO 105.

[00060] The antibody "9D7" is intended to mean an antibody consisting of the Light Chain SEQ ID NO 106 and Heavy Chain SEQ ID NO 107.

[00061] The antibody "9G11" is intended to mean an antibody consisting of the Light Chain SEQ ID NO 108 and Heavy Chain SEQ ID NO 109.

15 [00062] The antibody "L3" is intended to mean an antibody consisting of the Light Chain SEQ ID NO 112 and Heavy Chain SEQ ID NO 113.

[00063] The antibody "7A10" is intended to mean an antibody consisting of the Light Chain SEQ ID NO 114 and Heavy Chain SEQ ID NO 115.

[00064] The antibody "8D9" is intended to mean an antibody consisting of the Light 20 Chain SEQ ID NO 116 and Heavy Chain SEQ ID NO 117.

[00065] The antibody "9C12" is intended to mean an antibody consisting of the Light Chain SEQ ID NO 118 and Heavy Chain SEQ ID NO 119.

[00066] The antibody "7C4" is intended to mean an antibody consisting of the Light Chain SEQ ID NO 110 and Heavy Chain SEQ ID NO 111. 25 [00067] Unless otherwise specified herein, the numbering of amino acid residues in this region is according to IMGT, Sequences of Proteins of Immunological

Interest, 5th Ed. Public Health Service, National Institutes of Health, Be thesda, MD. (1991).

[00068] The above-mentioned antibodies can be used in in treating patients with Alzheimer's disease or patients with a taupahty such as Argyrophilic Grain Dis 5 ease (AGD), Psychosis, particularly Psychosis due to AD or Psychosis in pa tients with AD, psychiatric symptoms of patients with Lewy body dementia, Pro gressive Supranuclear Palsy (PSP), Frontotemporal dementia (FTD or variants thereof), TBI (traumatic brain injury, acute or chronic), Corticobasal Degenera tion (CBD), Picks Disease, Primary age-related tauopathy (PART), Neurofibril 10 lary tangle-predominant senile dementia, Dementia pugilistica, Chronic trau matic encephalopathy, stroke, stroke recovery, neurodegeneration in relation to Parkinson's disease, Parkinsonism linked to chromosome, Lytico-Bodig dis ease (Parkinson-dementia complex of Guam), Ganglioglioma and gangli ocytoma, Meningioangiomatosis, Postencephalitic parkinsonism, Subacute 15 sclerosing panencephalitis, Huntington's disease, lead encephalopathy, tuber ous sclerosis, Hallervorden-Spatz disease and lipofuscinosis. More typically, the taupathy is selected from the group consisting of Alzheimer's disease, Ar gyrophilic Grain Disease (AGD), Psychosis, particularly Psychosis due to AD or Psychosis in patients with AD, psychiatric symptoms of patients with Lewy 20 body dementia, Progressive Supranuclear Palsy (PSP), Frontotemporal de mentia (FTD or variants thereof), TBI (traumatic brain injury, acute or chronic), Corticobasal Degeneration (CBD), and Picks Disease.

[00069] An "anti-alpha-synuclein" antibody is an antibody which binds specifi cally to alpha-synuclein or an alpha-synuclein fragment. 25 [00070] The term "human antibody", as used herein, is intended to include anti bodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the invention may in clude amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site- specific mutagene 30 sis in vitro or during gene rearrangement or by somatic mutation in vivo).

[00071] The terms "monoclonal antibody" or "monoclonal antibody composition" as used herein refer to a preparation of antibody molecules of single molecu lar composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope. 5 [00072] The antibodies of the present invention, and their alpha-synuclein epitope-binding fragments will preferably be "humanized," particularly if em ployed for therapeutic purposes. The term "humanized" refer to a chimeric molecule, generally prepared using recombinant techniques, having an anti gen binding site derived from an immunoglobulin from a non-human species 10 and a remaining immunoglobulin structure based upon the structure and /or sequence of a human immunoglobulin. The antigen-binding site may com prise either complete non-human antibody variable domains fused to human constant domains, or only the complementarity determining regions (CDRs) of such variable domains grafted to appropriate human framework regions of 15 human variable domains. The framework residues of such humanized mole cules may be wild type (e.g., fully human) or they may be modified to contain one or more amino acid substitutions not found in the human antibody whose sequence has served as the basis for humanization. Humanization lessens or eliminates the likelihood that a constant region of the molecule will act as 20 an immunogen in human individuals, but the possibility of an immune re sponse to the foreign variable region remains (LoBuglio, A.F. et al. (1989) "Mouse/Human Chimeric Monoclonal Antibody In Man: Kinetics And Immune Response," Proc. Natl. Acad. Sci. (U.S.A.) 86:4220-4224). Another approach focuses not only on providing human-derived constant regions, but modifying 25 the variable regions as well so as to reshape them as closely as possible to human form. It is known that the variable regions of both heavy and light chains contain three complementarity- determining regions (CDRs) which vary in response to the antigens in question and determine binding capability, flanked by four framework regions (FRs) which are relatively conserved in a 30 given species and which putatively provide a scaffolding for the CDRs. When non-human antibodies are prepared with respect to a particular antigen, the variable regions can be "reshaped" or "humanized" by grafting CDRs derived from nonhuman antibody on the FRs present in the human antibody to be modified. Application of this approach to various antibodies has been re ported by Sato, K. et al. (1993) Cancer Res 53:851-856. Riechmann, L. et al. (1988) "Reshaping Human Antibodies for Therapy," Nature 332:323-327; Verhoeyen, M. et al. (1988) "ReshapingHuman Antibodies: Grafting An Anti 5 lysozyme Activity," Science 239:1534-1536; Kettleborough, C. A. et al. (1991) "Humanization Of A Mouse Monoclonal Antibody By CDR-Grafting: The Im portance Of Framework Residues On Loop Conformation," Protein Engineer ing 4:773-3783; Maeda, H. et al. (1991) "Construction Of Reshaped Human Antibodies With HIV-Neutralizing Activity," Human Antibodies Hybridoma 10 2:124-134; Gorman, S. D. et al. (1991) "Reshaping A Therapeutic CD4 Anti body," Proc. Nati. Acad. Sci. (U.S.A.) 88:4181-4185; Tempest, P.R. et al. (1991) "ReshapingA Human Monoclonal Antibody To Inhibit Human Respira tory Syncytial Virus Infection in vivo," Bio/Technology 9:266-271; Co, M. S. et al. (1991) "HumanizedAntibodies For Antiviral Therapy," Proc. Nat. Acad. 15 Sci. (U.S.A.) 88:2869-2873; Carter, P. et al. (1992) "Humanization Of An Anti p185her2 Antibody For Human Cancer Therapy," Proc. Nati. Acad. Sci. (U.S.A.) 89:4285-4289; and Co, M.S. et al. (1992) "Chimeric And Humanized Antibodies With Specificity For The CD33 Antigen," J. Immunol. 148:1149 1154. In some embodiments, humanized antibodies preserve all CDR se 20 quences (for example, a humanized mouse antibody which contains all six CDRs from the mouse antibodies). In other embodiments, humanized anti bodies have one or more CDRs (one, two, three, four, five, six) which are al tered with respect to the original antibody, which are also termed one or more CDRs "derived from" one or more CDRs from the original antibody. The abil 25 ity to humanize an antigen is well known (see, e.g., US Patents No. 5,225,539; 5,530,101; 5,585,089; 5,859,205; 6,407,213; 6,881,557).

[00073] As used herein, an antibody or an epitope-binding antigen-binding frag ment thereof is said to "immunospecifically" bind a region of another molecule (i.e., an epitope) if it reacts or associates more frequently, more rapidly, with 30 greater duration and/or with greater affinity or avidity with that epitope relative to alternative epitopes. It is also understood by reading this definition that, for example, an antibody or an epitope-binding antigen-binding fragment thereof that specifically binds to a first target may or may not specifically or preferen tially bind to a second target. As used herein, the term "binding" in the con text of the binding of an antibody to a predetermined antigen typically refers to binding with an affinity corresponding to a KD of about 10-7 M or less, such 5 as about 10-8 M or less, such as about 10-9 M or less when determined by for instance surface plasmon resonance (SPR) technology in a BAcore 3000 in strument using the antigen as the ligand and the antibody as the analyte, and binds to the predetermined antigen with an affinity corresponding to a KD that is at least ten-fold lower, such as at least 100 fold lower, for instance at least 10 1,000 fold lower, such as at least 10,000 fold lower, for instance at least 100,000 fold lower than its affinity for binding to a non-specific antigen (e.g., BSA, casein) other than the predetermined antigen or a closely-related anti gen. The amount with which the affinity is lower is dependent on the KD of the antibody, so that when the KD of the antibody is very low (that is, the anti 15 body is highly specific), then the amount with which the affinity for the antigen is lower than the affinity for a non-specific antigen may be at least 10,000 fold.

[00074] The term "kd" (sec -1 or 1/s), as used herein, refers to the dissociation rate constant of a particular antibody-antigen interaction. Said value is also 20 referred to as the koff value.

[00075] The term "ka" (M-1 x sec-1 or 1/M), as used herein, refers to the associ ation rate constant of a particular antibody-antigen interaction.

[00076] The term "KD" (M), as used herein, refers to the dissociation equilibrium constant of a particular antibody-antigen interaction. 25 [00077] The term "KA" (M-1 or 1/M), as used herein, refers to the association equilibrium constant of a particular antibody-antigen interaction and is ob tained by dividing the ka by the kd.

[00078] In some antibodies only part of a CDR, namely the subset of CDR resi dues required for binding, termed the SDRs, are needed to retain binding in a 30 humanized antibody. CDR residues not contacting antigen and not in the SDRs can be identified based on previous studies (for example residues H60 H65 in CDR H2 are often not required), from regions of Kabat CDRs lying outside Chothia hypervariable loops (see, Kabat et al. (1992) SEQUENCES OF

PROTEINS OF IMMUNOLOGICAL INTEREST, National Institutes of Health Publica tion No. 91-3242; Chothia, C. et al. (1987) "Canonical Structures For The Hy pervariable Regions OfImmunoglobulins," J. Mol. Biol. 196:901-917), by mo lecular modeling and/or empirically, or as described in Gonzales, N.R. et al. 5 (2004) "SDR Grafting Of A Murine Antibody Using Multiple Human Germline Templates To Minimize Its Immunogenicity," Mol. Immunol. 41:863-872. In such humanized antibodies at positions in which one or more donor CDR res idues is absent or in which an entire donor CR is omitted, the amino acid oc cupying the position can be an amino acid occupying the corresponding posi 10 tion (by Kabat numbering) in the acceptor antibody sequence. The number of such substitutions of acceptor for donor amino acids in the CDRs to include reflects a balance of competing considerations. Such substitutions are poten tially advantageous in decreasing the number of mouse amino acids in a hu manized antibody and consequently decreasing potential immunogenicity. 15 However, substitutions can also cause changes of affinity, and significant re ductions in affinity are preferably avoided. Positions for substitution within CDRs and amino acids to substitute can also be selected empirically.

[00079] The fact that a single amino acid alteration of a CDR residue can result in loss of functional binding (Rudikoff, S. etc. (1982) "Single Amino Acid Sub 20 stitution Altering Antigen-Binding Specificity," Proc. Natl. Acad. Sci. (USA) 79(6):1979-1983) provides a means for systematically identifying alternative functional CDR sequences. In one preferred method for obtaining such vari ant CDRs, a polynucleotide encoding the CDR is mutagenized (for example via random mutagenesis or by a site-directed method (e.g., polymerase 25 chain-mediated amplification with primers that encode the mutated locus)) to produce a CDR having a substituted amino acid residue. By comparing the identity of the relevant residue in the original (functional) CDR sequence to the identity of the substituted (non-functional) variant CDR sequence, the BLOSUM62.iij substitution score for that substitution can be identified. The 30 BLOSUM system provides a matrix of amino acid substitutions created by an alyzing a database of sequences for trusted alignments (Eddy, S.R. (2004) "Where Did The BLOSUM62 Alignment Score Matrix Come From?," Nature

Biotech. 22(8):1035-1036; Henikoff, J.G. (1992) "Amino acid substitution ma trices from protein blocks," Proc. Nat. Acad. Sci. (USA) 89:10915-10919; Karlin, S. et al. (1990) "Methods For Assessing The Statistical Significance Of Molecular Sequence Features By Using General Scoring Schemes," Proc. 5 Nat. Acad. Sci. (USA) 87:2264-2268; Altschul, S.F. (1991) "Amino Acid Sub stitution Matrices From An Information Theoretic Perspective,"J. Mol. Biol. 219, 555-565. Currently, the most advanced BLOSUM database is the BLOSUM62 database (BLOSUM62.iij). Table 1 presents the BLOSUM62.iij substitution scores (the higher the score the more conservative the substitu 10 tion and thus the more likely the substitution will not affect function). If an an tigen-binding fragment comprising the resultant CDR fails to bind to ROR1, for example, then the BLOSUM62.iij substitution score is deemed to be insuf ficiently conservative, and a new candidate substitution is selected and pro duced having a higher substitution score. Thus, for example, if the original 15 residue was glutamate (E), and the non-functional substitute residue was his tidine (H), then the BLOSUM62.iij substitution score will be 0, and more con servative changes (such as to aspartate, asparagine, glutamine, or lysine) are preferred.

Table 1 A R N D C Q E G H I L K M F P S T W Y V A +4 -1 -2 -2 0 -1 -1 0 -2 -1 -1 -1 -1 -2 -1 +1 0 -3 -2 0 R -1 +5 0 -2 -3 +1 0 -2 0 -3 -2 +2 -1 -3 -2 -1 -1 -3 -2 -3 N -2 0 +6 +1 -3 0 0 0 +1 -3 -3 0 -2 -3 -2 +1 0 -4 -2 -3 D -2 -2 +1 +6 -3 0 +2 -1 -1 -3 -4 -1 -3 -3 -1 0 -1 -4 -3 -3 C 0 -3 -3 -3 +9 -3 -4 -3 -3 -1 -1 -3 -1 -2 -3 -1 -1 -2 -2 -1 0 -1 +1 0 0 -3 +5 +2 -2 0 -3 -2 +1 0 -3 -1 0 -1 -2 -1 -2 E -1 0 0 +2 -4 +2 +5 -2 0 -3 -3 +1 -2 -3 -1 0 -1 -3 -2 -2 G 0 -2 0 -1 -3 -2 -2 +6 -2 -4 -4 -2 -3 -3 -2 0 -2 -2 -3 -3 H -2 0 +1 -1 -3 0 0 -2 +8 -3 -3 -1 -2 -1 -2 -1 -2 -2 +2 -3 1 -1 -3 -3 -3 -1 -3 -3 -4 -3 +4 +2 -3 +1 0 -3 -2 -1 -3 -1 +3 L -1 -2 -3 -4 -1 -2 -3 -4 -3 +2 +4 -2 +2 0 -3 -2 -1 -2 -1 +1 K -1 +2 0 -1 -3 +1 +1 -2 -1 -3 -2 +5 -1 -3 -1 0 -1 -3 -2 -2 M -1 -1 -2 -3 -1 0 -2 -3 -2 +1 +2 -1 +5 0 -2 -1 -1 -1 -1 +1 F -2 -3 -3 -3 -2 -3 -3 -3 -1 0 0 -3 0 +6 -4 -2-2 +1 +3 -1 P -1 -2 -2 -1 -3 -1 -1 -2 -2 -3 -3 -1 -2 -4 +7 -1 -1 -4 -3 -2 S +1 -1 +1 0 -1 0 0 0 -1 -2 -2 0 -1 -2 -1 +4 +1 -3 -2 -2 T 0 -1 0 -1 -1 -1 -1 -2 -2 -1 -1 -1 -1 -2 -1 +1 +5 -2 -2 0 W -3 -3 -4 -4 -2 -2 -3 -2 -2 -3 -2 -3 -1 +1 -4 -3 -2 +11 +2 -3 Y -2 -2 -2 -3 -2 -1 -2 -3 +2 -1 -1 -2 -1 +3 -3 -2 -2 +2 +7 -1 V 0 -3 -3 -3 -1 -2 -2 -3 -3 +3 +1 -2 +1 -1 -2 -2 0 -3 -1 +4

[00080] The invention thus contemplates the use of random mutagenesis to identify improved CDRs. In the context of the present invention, conservative substitutions may be defined by substitutions within the classes of amino ac 5 ids reflected in one or more of the following three tables:

Amino Acid Residue Classes For Conservative Substitutions: Table 2 Acidic Residues Asp (D) and Glu (E) Basic Residues Lys (K), Arg (R), and His (H) Hydrophilic Uncharged Residues Ser (S), Thr (T), Asn (N), and Gln (Q)

Aliphatic Uncharged Residues Cly (G), Ala (A), Val (V), Leu (L), and Ile (1) Non-polar Uncharged Residues Cys (C), Met (M), and Pro (P) Aromatic Residues Phe (F), Tyr (Y), and Trp (W)

Alternative Conservative Amino Acid Residue Substitution Classes: Table 3 1 A S T 2 D E 3 N Q 4 R K 5 I L M 6 F Y W Alternative Physical and Functional Classifications of Amino Acid Resi dues: 5 Table 4 Alcohol Group-Containing Residues S and T Aliphatic Residues I, L, V and M Cycloalkenyl-Associated Residues F, H, W and Y Hydrophobic Residues A, C, F, G, H, I, L, M, R, T, V, W and Y Negatively Charged Residues D and E Polar Residues C, D, E, H, K, N, Q, R, S and T Positively Charged Residues H, K and R Small Residues A, C, D, G, N, P, S, T and V Very Small Residues A, G and S Residues Involved In Turn Formation A, C, D, E, G, H, K, N, Q, R, S, P and T Flexible Residues Q, T, K, S, G, P, D, E and R

[00081] More conservative substitutions groupings include: valine-leucine-isoleu cine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, and asparagine glutamine.

[00082] Additional groups of amino acids may also be formulated using the prin 5 ciples described in, e.g., Creighton (1984) Proteins: Structure and Molecular Properties (2d Ed. 1993), W. H. Freeman and Company.

[00083] Phage display technology can alternatively be used to increase (or de crease) CDR affinity. This technology, referred to as affinity maturation, em ploys mutagenesis or "CDR walking" and re-selection uses the target antigen 10 or an antigenic antigen-binding fragment thereof to identify antibodies having CDRs that bind with higher (or lower) affinity to the antigen when compared with the initial or parental antibody (See, e.g. Glaser et al. (1992) J. Immunol ogy 149:3903). Mutagenizing entire codons rather than single nucleotides re sults in a semi-randomized repertoire of amino acid mutations. Libraries can 15 be constructed consisting of a pool of variant clones each of which differs by a single amino acid alteration in a single CDR and which contain variants rep resenting each possible amino acid substitution for each CDR residue. Mu tants with increased (or decreased) binding affinity for the antigen can be screened by contacting the immobilized mutants with labelled antigen. Any 20 screening method known in the art can be used to identify mutant antibodies with increased or decreased affinity to the antigen (e.g., ELISA) (See Wu et al. 1998, Proc. Natl. Acad. Sci. (U.S.A.) 95:6037; Yelton et al., 1995, J. Immu nology 155:1994). CDR walking which randomizes the Light Chain may be used possible (see, Schier et al., 1996, J. Mol. Bio. 263:551). 25 [00084] Methods for accomplishing such affinity maturation are described for ex ample in: Krause, J.C. et al. (2011) "An Insertion Mutation That Distorts Anti body Binding Site Architecture Enhances Function Of A Human Antibody," MBio. 2(1) pii: e00345-10. doi: 10.1128/mBio.00345-10; Kuan, C.T. et al. (2010) "Affinity-Matured Anti-Glycoprotein NMB Recombinant Immunotoxins 30 Targeting Malignant Gliomas And Melanomas," I nt. J. Cancer 10.1002/ijc.25645; Hackel, B.J. et al. (2010) "Stability And CDR Composition Biases Enrich Binder Functionality Landscapes," J. Mol. Biol. 401(1):84-96; Montgomery, D.L. et al. (2009) "Affinity Maturation And Characterization Of A

Human Monoclonal Antibody Against HIV-1 gp41," MAbs 1(5):462-474; Gustchina, E. et al. (2009) "Affinity Maturation By Targeted Diversification Of The CDR-H2 Loop Of A Monoclonal Fab Derived From A Synthetic Naive Hu man Antibody Library And Directed Against The Internal Trimeric Coiled-Coil 5 Of Gp41 Yields A Set Of Fabs With Improved HIV-1 Neutralization Potency And Breadth," Virology 393(1):112-119; Finlay, W.J. et al. (2009) "Affinity Maturation Of A Humanized Rat Antibody For Anti-RAGE Therapy: Compre hensive Mutagenesis Reveals A High Level Of Mutational Plasticity Both In side And Outside The Complementarity-Determining Regions," J. Mol. Biol. 10 388(3):541-558; Bostrom, J. et al. (2009) "Improving Antibody Binding Affinity And Specificity For Therapeutic Development," Methods Mol. Biol. 525:353 376; Steidl, S. et al. (2008) "In Vitro Affinity Maturation Of Human GM-CSF Antibodies By Targeted CDR-Diversification," Mol. Immunol. 46(1):135-144; and Barderas, R. et al. (2008) "Affinity Maturation Of Antibodies Assisted By 15 In Silico Modeling," Proc. Natl. Acad. Sci. (USA) 105(26):9029-9034.

[00085] Thus, the sequence of CDR variants of encompassed antibodies or their epitope-binding fragments may differ from the sequence of the CDR of the parent antibody through substitutions; for instance substituted 4 amino acid residue, 3 amino acid residue, 2 amino acid residue or 1 of the amino acid 20 residues. According to an embodiment of the invention it is furthermore envis aged that the amino acids in the CDR regions may be substituted with con servative substitutions, as defined in the below 3 tables. For example, the acidic residue Asp can be substituted with Glu without substantially affecting the binding characteristic of the antibody. 25 [00086] The term "epitope" means an antigenic determinant capable of immuno specific binding to an antibody. Epitopes usually consist of surface groupings of molecules such as amino acids or sugar side chains and usually have spe cific three dimensional structural characteristics, as well as specific charge characteristics. Conformational and nonconformational epitopes are distin 30 guished in that the binding to the former, but not the latter, is lost in the pres ence of denaturing solvents. The epitope may comprise amino acid residues directly involved in the binding (also called immunodominant component of the epitope) and other amino acid residues, which are not directly involved in the binding, such as amino acid residues which are effectively blocked by the specifically antigen binding peptide (in other words, the amino acid residue is within the footprint of the specifically antigen binding peptide).

[00087] The term "transgenic non-human animal" refers to a non-human animal 5 having a genome comprising one or more human heavy and/or light chain transgenes or transchromosomes (either integrated or non-integrated into the animal's natural genomic DNA) and which is capable of expressing fully hu man antibodies. For example, a transgenic mouse can have a human light chain transgene and either a human heavy chain transgene or human heavy 10 chain transchromosome, such that the mouse produces human anti-alpha synuclein antibody when immunized with alpha-synuclein antigen and/or cells expressing alpha-synuclein. The human heavy chain transgene may be inte grated into the chromosomal DNA of the mouse, as is the case for transgenic mice, for instance HuMAb mice, such as HCo7 or HCol2 mice, or the human 15 heavy chain transgene may be maintained extrachromosomally, as is the case for transchromosomal KM mice as described in W002/43478. Such transgenic and transchromosomal mice (collectively referred to herein as "transgenic mice") are capable of producing multiple isotypes of human mon oclonal antibodies to a given antigen (such as IgG, IgA, IgM, IgD and/or IgE) 20 by undergoing V-D-J recombination and isotype switching.

[00088] Transgenic, nonhuman animal can also be used for production of anti bodies against a specific antigen by introducing genes encoding such specific antibody, for example by operatively linking the genes to a gene which is ex pressed in the milk of the animal. 25 [00089] The term "treatment" or "treating" as used herein means ameliorating, slowing or reversing the progress or severity of a disease or disorder, or ame liorating, slowing or reversing one or more symptoms or side effects of such disease or disorder. For purposes of this invention, "treatment" or "treating" further means an approach for obtaining beneficial or desired clinical results, 30 where "beneficial or desired clinical results" include, without limitation, allevia tion of a symptom, diminishment of the extent of a disorder or disease, stabi lized (i.e., not worsening) disease or disorder state, delay or slowing of the progression a disease or disorder state, amelioration or palliation of a disease or disorder state, and remission of a disease or disorder, whether partial or total, detectable or undetectable.

[00090] An "effective amount," when applied to an antibody of the invention, re fers to an amount sufficient, at dosages and for periods of time necessary, to 5 achieve an intended biological effect or a desired therapeutic result including, without limitation, clinical results. The phrase "therapeutically effective amount" when applied to an antibody of the invention is intended to denote an amount of the antibody that is sufficient to ameliorate, palliate, stabilize, re verse, slow or delay the progression of a disorder or disease state, or of a 10 symptom of the disorder or disease. In an embodiment, the method of the present invention provides for administration of the antibody in combinations with other compounds. In such instances, the "effective amount" is the amount of the combination sufficient to cause the intended biological effect.

[00091] A therapeutically effective amount of an anti-alpha-synuclein antibody 15 may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the anti-alpha-synuclein antibody to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the antibody or antibody por tion are outweighed by the therapeutically beneficial effects. 20 [00092] As indicated above, the present invention particularly relates to a mono clonal antibody capable of immunospecifically binding to an epitope within amino acids 110-140 of human alpha-synuclein. In one embodiment the anti body is capable of competing with the antibody GM37 for binding the 112-117 epitope of alpha-synuclein. In another embodiment the antibody is capable of 25 competing with the antibody GM285 for binding the 112-115 epitope of al pha-synuclein, In another embodiment the antibody is capable of competing with the antibody GM63 for binding the 126-138 epitope of alpha-synuclein. In another embodiment the antibody is capable of competing with the antibody 2E6 for binding the 126-140 epitope of alpha-synuclein. In yet another em 30 bodiment the antibody is capable of competing with the antibody 9E4 for bind ing the 118-126 epitope of alpha-synuclein.

[00093] The antibody is preferably a human or humanized antibody.

[00094] The antibodies of the invention is further defined in the claims

[00095] The present invention also provides a method of reducing Tau tangle formation in a patient, comprising administering to the patient in need of such treatment, a therapeutically effective amount of an antibody of the invention.

[00096] Further the antibodies may be in a composition together with a pharma 5 ceutically acceptable carrier, diluent and/or stabilizer. The antibodies of the invention may be used in therapy. In particular the antibodies of the invention may be used in treating a taupathy. Typically, the taupathy is selected from the group consisting of Alzheimer's disease, Argyrophilic Grain Disease (AGD), Psychosis, particularly Psychosis due to AD or Psychosis in patients 10 with AD, psychiatric symptoms of patients with Lewy body dementia, Progres sive Supranuclear Palsy (PSP), Frontotemporal dementia (FTD or variants thereof), TBI (traumatic brain injury, acute or chronic), Corticobasal Degener ation (CBD), Picks Disease, Primary age-related tauopathy (PART), Neurofi brillary tangle-predominant senile dementia, Dementia pugilistica, Chronic 15 traumatic encephalopathy, stroke, stroke recovery, neurodegeneration in rela tion to Parkinson's disease, Parkinsonism linked to chromosome, Lytico Bodig disease (Parkinson-dementia complex of Guam), Ganglioglioma and gangliocytoma, Meningioangiomatosis, Postencephalitic parkinsonism, Sub acute sclerosing panencephalitis, Huntington's disease, lead encephalopathy, 20 tuberous sclerosis, Hallervorden-Spatz disease and lipofuscinosis. More typi cally, the taupathy is selected from the group consisting of Alzheimer's dis ease, Argyrophilic Grain Disease (AGD), Psychosis, particularly Psychosis due to AD or Psychosis in patients with AD, psychiatric symptoms of patients with Lewy body dementia, Progressive Supranuclear Palsy (PSP), Frontotem 25 poral dementia (FTD or variants thereof), TBI (traumatic brain injury, acute or chronic), Corticobasal Degeneration (CBD), and Picks Disease. In particular, the tauopathies may be selected from Alzheimer's disease, Argyrophilic Grain Disease (AGD), Psychosis due to AD or Psychosis in patients with AD, and psychiatric symptoms of patients with Lewy body dementia. 30 [00097] The treatment may be chronic and the patient may be treated at least 2 weeks, such as at least for 1 month, 6, months, 1 year or more.

[00098] The antibodies of the present invention may for example be monoclonal antibodies produced by the hybridoma method first described by Kohler et al.,

Nature 256, 495 (1975), or may be produced by recombinant DNA methods. Monoclonal antibodies may also be isolated from phage antibody libraries us ing the techniques described in, for example, Clackson et al., Nature 352, 624-628 (1991) and Marks et al., J. Mol. Biol. 222, 581-597 (1991). Monoclo 5 nal antibodies may be obtained from any suitable source. Thus, for example, monoclonal antibodies may be obtained from hybridomas prepared from mu rine splenic B lymphocyte cells obtained from mice immunized with an anti gen of interest, for instance, in the form of cells expressing the antigen on the surface, or a nucleic acid encoding an antigen of interest. Monoclonal anti 10 bodies may also be obtained from hybridomas derived from antibody-ex pressing cells of immunized humans or from non-human mammals such as rats, rabbits, dogs, sheep, goats, primates, etc.

[00099] In one embodiment, the antibody of the invention is a human antibody. Human monoclonal antibodies directed against alpha-synuclein may be gen 15 erated using transgenic or transchromosomal mice carrying parts of the hu man immune system rather than the mouse system. Such transgenic and transchromosomic mice include mice referred to herein as HuMAb mice and KM mice, respectively, and are collectively referred to herein as "transgenic mice". 20 [000100] The HuMAb mouse contains a human immunoglobulin gene mini locus that encodes unrearranged human heavy variable and constant (p and Y) and light variable and constant (K) chain immunoglobulin sequences, to gether with targeted mutations that inactivate the endogenous p and K chain loci (Lonberg, N. et al., Nature 368, 856-859 (1994)). Accordingly, such mice 25 exhibit reduced expression of mouse IgM or IgK and in response to immun ization, the introduced human heavy and light chain transgenes, undergo class switching and somatic mutation to generate high affinity human IgG, i monoclonal antibodies (Lonberg, N. et al. (1994), supra; reviewed in Lonberg, N., Handbook of Experimental Pharmacology 113, 49-101 (1994) , Lonberg, 30 N. and Huszar, D., Intern. Rev. Immunol. Vol. 13 65-93 (1995) and Harding, F. and Lonberg, N., Ann. N. Y. Acad. Sci 764 536-546 (1995)). The prepara tion of HuMAb mice is described in detail in Taylor, L. et al., Nucleic Acids Research 20, 6287-6295 (1992), Chen, J. et al., International Immunology 5,

647-656 (1993), Tuaillon et al., J. Immunol. 152, 2912-2920 (1994), Taylor, L. et al., International Immunology 6, 579-591 (1994), Fishwild, D. et al., Nature Biotechnology 14, 845-851 (1996). See also US 5,545,806, US 5,569,825, US 5,625,126, US 5,633,425, US 5,789,650, US 5,877,397, US 5,661,016, 5 US 5,814,318, US 5,874,299, US 5,770,429, US 5,545,807, WO 98/24884, WO 94/25585, WO 93/1227, WO 92/22645, WO 92/03918 and WO 01/09187.

[000101] The HCo7 mice have a JKD disruption in their endogenous light chain (kappa) genes (as described in Chen et al., EMBO J. 12, 821-830 10 (1993)), a CMD disruption in their endogenous heavy chain genes (as de scribed in Example 1 of WO 01/14424), a KCo5 human kappa light chain transgene (as described in Fishwild et al., Nature Biotechnology 14, 845-851 (1996)), and a HCo7 human heavy chain transgene (as described in US 5,770,429). 15 [000102] The HCol2 mice have a JKD disruption in their endogenous light chain (kappa) genes (as described in Chen et al., EMBO J. 12, 821-830 (1993)), a CMD disruption in their endogenous heavy chain genes (as de scribed in Example 1 of WO 01/14424), a KCo5 human kappa light chain transgene (as described in Fishwild et al., Nature Biotechnology 14, 845-851 20 (1996)), and a HCol2 human heavy chain transgene (as described in Exam ple 2 of WO 01/14424).

[000103] In the KM mouse strain, the endogenous mouse kappa light chain gene has been homozygously disrupted as described in Chen et al., EMBO J. 12, 811-820 (1993) and the endogenous mouse heavy chain gene has been 25 homozygously disrupted as described in Example 1 of WO 01/09187. This mouse strain carries a human kappa light chain transgene, KCo5, as de scribed in Fishwild et al., Nature Biotechnology 14, 845-851 (1996). This mouse strain also carries a human heavy chain transchromosome composed of chromosome 14 fragment hCF (SC20) as described in WO 02/43478. 30 [000104] Splenocytes from these transgenic mice may be used to generate hybridomas that secrete human monoclonal antibodies according to well known techniques. Human monoclonal of the present invention, or antibodies of the present invention originating from other species may also be generated transgenically through the generation of another non-human mammal or plant that is transgenic for the immunoglobulin heavy and light chain sequences of interest and production of the antibody in a recoverable form therefrom. In connection with the transgenic production in mammals, antibodies may be 5 produced in, and recovered from, the milk of goats, cows, or other mammals. See for instance US 5,827,690, US 5,756,687, US 5,750,172 and US 5,741,957.

[000105] The antibody of the invention may be of any isotype. The choice of isotype typically will be guided by the desired effector functions, such as 10 ADCC induction. Exemplary isotypes are IgGI, IgG2, IgG3, and IgG4. Either of the human light chain constant regions, kappa or lambda, may be used. If desired, the class of an anti-alpha-synuclein antibody of the present invention may be switched by known methods. For example, an antibody of the present invention that was originally IgM may be class switched to an IgG antibody of 15 the present invention. Further, class switching techniques may be used to convert one IgG subclass to another, for instance from IgGI to IgG2. Thus, the effector function of the antibodies of the present invention may be changed by isotype switching to, e.g., an IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM antibody for various therapeutic uses. In one embodiment an anti 20 body of the present invention is an IgG1 antibody, for instance an IgG1, K.

[000106] In one embodiment, the antibody of the invention is a full-length antibody, preferably an IgG antibody, in particular an IgG1, K antibody. In an other embodiment, the antibody of the invention is an antibody fragment or a single-chain antibody. 25 [000107] Antibodies fragments may e.g. be obtained by fragmentation using conventional techniques, and the fragments screened for utility in the same manner as described herein for whole antibodies. For example, F(ab')2 frag ments may be generated by treating antibody with pepsin. The resulting F(ab')2 fragment may be treated to reduce disulfide bridges to produce Fab' 30 fragments. Fab fragments may be obtained by treating an IgG antibody with papain; Fab'fragments may be obtained with pepsin digestion of IgG anti body. An F(ab') fragment may also be produced by binding Fab'described below via a thioether bond or a disulfide bond. A Fab'fragment is an antibody fragment obtained by cutting a disulfide bond of the hinge region of the F(ab')2. A Fab'fragment may be obtained by treating an F(ab')2 fragment with a reducing agent, such as dithiothreitol. Antibody fragment may also be generated by expression of nucleic acids encoding such fragments in recom 5 binant cells (see for instance Evans et al., J. Immunol. Meth. 184, 123-38 (1995)). For example, a chimeric gene encoding a portion of an F(ab')2 frag ment could include DNA sequences encoding the CH1 domain and hinge re gion of the H chain, followed by a translational stop codon to yield such a truncated antibody fragment molecule. 10 [000108] In one embodiment, the anti-alpha-synuclein antibody is a mono valent antibody, preferably a monovalent antibody as described in W02007059782 (which is incorporated herein by reference in its entirety) having a deletion of the hinge region. Accordingly, in one embodiment, the antibody is a monovalent antibody, wherein said anti-alpha-synuclein anti 15 body is constructed by a method comprising: i) providing a nucleic acid con struct encoding the light chain of said monovalent antibody, said construct comprising a nucleotide sequence encoding the VL region of a selected anti gen specific anti-alpha-synuclein antibody and a nucleotide sequence encod ing the constant CL region of an Ig, wherein said nucleotide sequence encod 20 ing the VL region of a selected antigen specific antibody and said nucleotide sequence encoding the CL region of an Ig are operably linked together, and wherein, in case of an IgG1 subtype, the nucleotide sequence encoding the CL region has been modified such that the CL region does not contain any amino acids capable of forming disulfide bonds or covalent bonds with other 25 peptides comprising an identical amino acid sequence of the CL region in the presence of polyclonal human IgG or when administered to an animal or hu man being; ii) providing a nucleic acid construct encoding the heavy chain of said monovalent antibody, said construct comprising a nucleotide sequence encoding the VH region of a selected antigen specific antibody and a nucleo 30 tide sequence encoding a constant CH region of a human Ig, wherein the nu cleotide sequence encoding the CH region has been modified such that the region corresponding to the hinge region and, as required by the Ig subtype, other regions of the CH region, such as the CH3 region, does not comprise any amino acid residues which participate in the formation of disulphide bonds or covalent or stable non-covalent inter-heavy chain bonds with other peptides comprising an identical amino acid sequence of the CH region of the human Ig in the presence of polyclonal human IgG or when administered to 5 an animal human being, wherein said nucleotide sequence encoding the VH region of a selected antigen specific antibody and said nucleotide sequence encoding the CH region of said Ig are operably linked together; iii) providing a cell expression system for producing said monovalent antibody; iv) producing said monovalent antibody by co-expressing the nucleic acid constructs of (i) 10 and (ii) in cells of the cell expression system of (iii).

[000109] Similarly, in one embodiment, the anti- alpha-synuclein antibody is a monovalent antibody, which comprises: (i) a variable region of an antibody of the invention as described herein or an anti gen binding part of the said region, and 15 (ii) a CH region of an immunoglobulin or a antigen-binding fragment thereof com prising the CH2 and CH3 regions, wherein the CH region or antigen-binding frag ment thereof has been modified such that the region corresponding to the hinge region and, if the immunoglobulin is not an IgG4 subtype, other regions of the CH region, such as the CH3 region, do not comprise any amino acid residues, which 20 are capable of forming disulfide bonds with an identical CH region or other cova lent or stable non-covalent inter-heavy chain bonds with an identical CH region in the presence of polyclonal human IgG.

[000110] In a further embodiment, the heavy chain of the monovalent anti alpha-synuclein antibody has been modified such that the entire hinge has 25 been deleted.

[000111] In another further embodiment, the sequence of said monovalent antibody has been modified so that it does not comprise any acceptor sites for N-linked glycosylation.

[000112] Anti-alpha-synuclein antibodies of the invention also include single 30 chain antibodies. Single chain antibodies are peptides in which the heavy and light chain Fv regions are connected. In one embodiment, the present inven tion provides a single-chain Fv (scFv) wherein the heavy and light chains in the Fv of an anti-alpha-synuclein antibody of the present invention are joined with a flexible peptide linker (typically of about 10, 12, 15 or more amino acid residues) in a single peptide chain. Methods of producing such antibodies are described in for instance US 4,946,778, Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, 5 New York, pp. 269-315 (1994), Bird et al., Science 242, 423-426 (1988), Hu ston et al., PNAS USA 85, 5879-5883 (1988) and McCafferty et al., Nature 348, 552-554 (1990). The single chain antibody may be monovalent, if only a single VH and VL are used, bivalent, if two VH and VL are used, or polyva lent, if more than two VH and VL are used. 10 [000113] In general, anti-alpha-synuclein antibodies described herein may be modified by inclusion of any suitable number of modified amino acids and/or associations with such conjugated substituents. Suitability in this con text is generally determined by the ability to at least substantially retain alpha synuclein selectivity and/or the anti-alpha-synuclein specificity associated 15 with the non-derivatized parent anti-alpha-synuclein antibody. The inclusion of one or more modified amino acids may be advantageous in, for example, increasing polypeptide serum half-life, reducing polypeptide antigenicity, or in creasing polypeptide storage stability. Amino acid(s) are modified, for exam ple, co-translationally or post-translationally during recombinant production 20 (e.g., N-linked glycosylation at N-X-S/T motifs during expression in mamma lian cells) or modified by synthetic means. Non-limiting examples of a modi fied amino acid include a glycosylated amino acid, a sulfated amino acid, a prenylated (e. g., farnesylated, geranylgeranylated) amino acid, an acetylated amino acid, an acylated amino acid, a PEGylated amino acid, a biotinylated 25 amino acid, a carboxylated amino acid, a phosphorylated amino acid, and the like. References adequate to guide one of skill in the modification of amino acids are replete throughout the literature. Example protocols are found in Walker (1998) Protein Protocols On CD-Rom, Humana Press, Totowa, NJ. The modified amino acid may, for instance, be selected from a glycosylated 30 amino acid, a PEGylated amino acid, a farnesylated amino acid, an acety lated amino acid, a biotinylated amino acid, an amino acid conjugated to a li pid moiety, or an amino acid conjugated to an organic derivatizing agent.

[000114] Anti-alpha-synuclein antibodies may also be chemically modified by covalent conjugation to a polymer to for instance increase their circulating half-life. Exemplary polymers, and methods to attach them to peptides, are il lustrated in for instance US 4,766,106, US 4,179,337, US 4,495,285 and US 5 4,609,546. Additional illustrative polymers include polyoxyethylated polyols and polyethylene glycol (PEG) (e.g., a PEG with a molecular weight of be tween about 1,000 and about 40,000, such as between about 2,000 and about 20,000, e.g., about 3,000-12,000 g/mol).

[000115] In a aspect, the invention relates to a pharmaceutical composition 10 comprising: - an anti-alpha-synuclein antibody as defined herein, and - a pharmaceutically-acceptable carrier.

[000116] The pharmaceutical compositions may be formulated with pharma ceutically acceptable carriers or diluents as well as any other known adju 15 vants and excipients in accordance with conventional techniques such as those disclosed in Remington : The Science and Practice of Pharmacy, 21th Edition, Gennaro, Ed., Mack Publishing Co., Easton, PA, 2005.

[000117] The pharmaceutically acceptable carriers or diluents as well as any other known adjuvants and excipients should be suitable for the chosen 20 compound of the present invention and the chosen mode of administration. Suitability for carriers and other components of pharmaceutical compositions is determined based on the lack of significant negative impact on the desired biological properties of the chosen compound or pharmaceutical composition of the present invention (e.g., less than a substantial impact (10% or less rel 25 ative inhibition, 5% or less relative inhibition, etc.)) on antigen binding.

[000118] A pharmaceutical composition of the present invention may also include diluents, fillers, salts, buffers, detergents (e.g., a nonionic detergent, such as Tween-20 or Tween- 80), stabilizers (e.g., sugars or protein-free amino acids), preservatives, tissue fixatives, solubilizers, and/or other materi 30 als suitable for inclusion in a pharmaceutical composition. The diluent is se lected to not to affect the biological activity of the combination. Examples of such diluents are distilled water, physiological phosphate-buffered saline, Ringer's solutions, dextrose solution, and Hank's solution. In addition, the pharmaceutical composition or formulation may also include other carriers, or non-toxic, nontherapeutic, non-immunogenic stabilizers and the like. The compositions may also include large, slowly metabolized macromolecules, such as proteins, polysaccharides like chitosan, polylactic acids, polyglycolic 5 acids and copolymers (e.g., latex functionalized sepharose, agarose, cellu lose, and the like), polymeric amino acids, amino acid copolymers, and lipid aggregates (e.g., oil droplets or liposomes).

[000119] The actual dosage levels of the active ingredients in the pharma ceutical compositions of the present invention may be varied so as to obtain 10 an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of ad ministration. The selected dosage level will depend upon a variety of pharma cokinetic factors including the activity of the particular compositions of the present invention employed, or the amide thereof, the route of administration, 15 the time of administration, the rate of excretion of the particular compound be ing employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the pa tient being treated, and like factors well known in the medical arts. 20 [000120] The pharmaceutical composition may be administered by any suit able route and mode, including: parenteral, topical, oral or intranasal means for prophylactic and/or therapeutic treatment. In one embodiment, a pharma ceutical composition of the present invention is administered parenterally. The phrases "parenteral administration" and "administered parenterally" as 25 used herein means modes of administration other than enteral and topical ad ministration, usually by injection, and include epidermal, intravenous, intra muscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, in tradermal, intraperitoneal, intratendinous, transtracheal, subcutaneous, sub cuticular, intraarticular, subcapsular, subarachnoid, intraspinal, intracranial, 30 intrathoracic, epidural and intrasternal injection and infusion.

[000121] Additional suitable routes of administering a compound of the pre sent invention in vivo and in vitro are well known in the art and may be se lected by those of ordinary skill in the art.

[000122] In one embodiment that pharmaceutical composition is adminis tered by intravenous or subcutaneous injection or infusion.

[000123] Pharmaceutically acceptable carriers include any and all suitable solvents, dispersion media, coatings, antibacterial and antifungal agents, iso 5 tonicity agents, antioxidants and absorption delaying agents, and the like that are physiologically compatible with a compound of the present invention.

[000124] Examples of suitable aqueous and nonaqueous carriers which may be employed in the pharmaceutical compositions of the present inven tion include water, saline, phosphate buffered saline, ethanol, dextrose, poly 10 ols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, corn oil, peanut oil, cottonseed oil, and sesame oil, carboxymethyl cellulose colloidal solutions, tragacanth gum and injectable organic esters, such as ethyl oleate, and/or various buffers. Other carriers are well known in the pharmaceutical arts. 15 [000125] Pharmaceutically acceptable carriers include sterile aqueous solu tions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. The use of such media and agents for pharmaceutically active substances is known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, 20 use thereof in the pharmaceutical compositions of the present invention is contemplated.

[000126] Pharmaceutical compositions of the present invention may also comprise pharmaceutically acceptable antioxidants for instance (1) water sol uble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bi 25 sulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble anti oxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), bu tylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha- tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like. 30 [000127] Pharmaceutical compositions of the present invention may also comprise isotonicity agents, such as sugars, polyalcohols, such as mannitol, sorbitol, glycerol or sodium chloride in the compositions.

[000128] The pharmaceutical compositions of the present invention may also contain one or more adjuvants appropriate for the chosen route of ad ministration such as preservatives, wetting agents, emulsifying agents, dis persing agents, preservatives or buffers, which may enhance the shelf life or 5 effectiveness of the pharmaceutical composition. The compounds of the pre sent invention may be prepared with carriers that will protect the compound against rapid release, such as a controlled release formulation, including im plants, transdermal patches, and microencapsulated delivery systems. Such carriers may include gelatin, glyceryl monostearate, glyceryl distearate, bio 10 degradable, biocompatible polymers such as ethylene vinyl acetate, polyan hydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid alone or with a wax, or other materials well known in the art. Methods for the preparation of such formulations are generally known to those skilled in the art. See, e.g., Sustained and Controlled Release Drug Delivery Systems, J. 15 R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

[000129] In one embodiment, the compounds of the present invention may be formulated to ensure proper distribution in vivo. Pharmaceutically accepta ble carriers for parenteral administration include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile 20 injectable solutions or dispersion. The use of such media and agents for phar maceutically active substances is known in the art. Except insofar as any con ventional media or agent is incompatible with the active compound, use thereof in the pharmaceutical compositions of the present invention is con templated. Supplementary active compounds may also be incorporated into 25 the compositions.

[000130] Pharmaceutical compositions for injection must typically be sterile and stable under the conditions of manufacture and storage. The composition may be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration. The carrier may be a aqueous 30 or nonaqueous solvent or dispersion medium containing for instance water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. The proper fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as glycerol, mannitol, sorbitol, 5 or sodium chloride in the composition. Prolonged absorption of the injectable compositions may be brought about by including in the composition an agent that delays absorption, for example, monostearate salts and gelatin. Sterile injectable solutions may be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of in 10 gredients e.g. as enumerated above, as required, followed by sterilization mi crofiltration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients e.g. from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, examples 15 of methods of preparation are vacuum drying and freeze-drying (lyophiliza tion) that yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[000131] Sterile injectable solutions may be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or 20 a combination of ingredients enumerated above, as required, followed by sterilization microfiltration. Generally, dispersions are prepared by incorporat ing the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, 25 examples of methods of preparation are vacuum drying and freeze-drying (ly ophilization) that yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[000132] Dosage regimens in the above methods of treatment and uses are adjusted to provide the optimum desired response (e.g., a therapeutic re 30 sponse). For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally re duced or increased as indicated by the exigencies of the therapeutic situation. Parenteral compositions may be formulated in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein re fers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the re 5 quired pharmaceutical carrier. The specification for the dosage unit forms of the present invention are dictated by and directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals. 10 [000133] The effective dosages and the dosage regimens for the anti alpha synuclein antibodies depend on the disease or condition to be treated and may be determined by the persons skilled in the art. On any given day that a dosage is given, the dosage may range from about 0.0001 to about 100 mg/kg, and more usually from about 0.01 to about 5 mg/kg, of the host body 15 weight. For example, dosages can be 1 mg/kg body weight or 10 mg/kg body weight or within the range of 1-10 mg/kg body weight. Exemplary dosages thus include: from about 0.1 to about 10 mg/kg/body weight, from about 0.1 to about 5 mg/kg/body weight, from about 0.1 to about 2 mg/kg/body weight, from about 0.1 to about 1 mg/kg/body weight, for instance about 0.15 20 mg/kg/body weight, about 0.2 mg/kg/body weight, about 0.5 mg/kg/body weight, about 1 mg/kg/body weight, about 1.5 mg/kg/body weight, about 2 mg/kg/body weight, about 5 mg/kg/body weight, or about 10 mg/kg/body weight.

[000134] A physician or veterinarian having ordinary skill in the art may 25 readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the anti-alpha-synuclein antibody employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is 30 achieved. In general, a suitable daily dose of a composition of the present in vention will be that amount of the compound which is the lowest dose effec tive to produce a therapeutic effect. Such an effective dose will generally de pend upon the factors described above. Administration may e.g. be intrave nous, intramuscular, intraperitoneal, or subcutaneous, and for instance ad ministered proximal to the site of the target. If desired, the effective daily dose of a pharmaceutical composition may be administered as two, three, four, 5 five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. While it is possible for a compound of the present invention to be administered alone, it is preferable to administer the compound as a pharmaceutical composition as described above. 10

Sequence Listing

SEQ ID NO:1 GM37 CDR 1 Heavy Chain SEQ ID NO:2 GM37 CDR 2 Heavy Chain SEQ ID NO:3 GM37 CDR 3 Heavy Chain SEQ ID NO:4 GM37 CDR 1 Light Chain SEQ ID NO:5 GM37 CDR 2 Light Chain SEQ ID NO:6 GM37 CDR 3 Light Chain SEQ ID NO:7 GM37 Heavy Chain Variable Domain SEQ ID NO:8 GM37 Light Chain Variable Domain SEQ ID NO:9 Epitope 112-117 of Human Alpha-Synuclein SEQ ID NO:10 Human Alpha-Synuclein SEQ ID NO:11 A-Syn-AAKK-BAP SEQ ID NO:12 A-Syn-BAAK-BAP SEQ ID NO:13 A-Syn-BBAA-BAP SEQ ID NO:14 A-Syn-BBKK-BAP SEQ ID NO:15 A-Syn-120-140_Del-BAP SEQ ID NO:16 Residues 1-119 of Human Alpha-Synuclein SEQ ID NO:17 Kappa Light Chain Constant domain SEQ ID NO:18 IgG1 Heavy Chain Constant domain SEQ ID NO:19 GM285 Epitope 112-115 SEQ ID NO:20 GM285CDR 1 Heavy Chain SEQ ID NO:21 GM285 CDR 2 Heavy Chain SEQ ID NO:22 GM285 CDR 3 Heavy Chain SEQ ID NO:23 GM285 CDR 1 Light Chain SEQ ID NO:24 GM285 CDR 2 Light Chain SEQ ID NO:25 GM285 CDR 3 Light Chain SEQ ID NO:26 GM285 Heavy Chain Variable Domain SEQ ID NO:27 GM285 Light Chain Variable Domain SEQ ID NO:28 GM285 IgG1 Heavy Chain Constant domain SEQ ID NO:29 GM285 Kappa Light Chain Constant domain SEQ ID NO:30 GM37 Variant 1 Heavy Chain Variable Domain SEQ ID NO:31 GM37 Variant 2 Heavy Chain Variable Domain SEQ ID NO:32 GM37 Variant 3 Heavy Chain Variable Domain SEQ ID NO:33 GM37 Variant 1 Heavy Chain CDR 2 SEQ ID NO:34 GM37 Variant 2 Heavy Chain CDR 2 SEQ ID NO:35 GM37 Variant 3 Heavy Chain CDR 2 SEQ ID NO:36 9E4 Binding Epitope SEQ ID NO:37 Human Beta-Synuclein SEQ ID NO:38 Human Gamma-Synuclein SEQ ID NO:39 Alpha-Synuclein Ortholog for Cynomolgus Monkey SEQ ID NO:40 Alpha-Synuclein Ortholog for Rat SEQ ID NO:41 Alpha-Synuclein Ortholog for Mouse SEQ ID NO:42 9E4 VH SEQ ID NO:43 9E4 VL SEQ ID NO:44 9E4 CDR 1 Heavy Chain

SEQ ID NO:45 9E4 CDR 2 Heavy Chain SEQ ID NO:46 9E4 CDR 3 Heavy Chain SEQ ID NO:47 9E4 CDR 1 Light Chain SEQ ID NO:48 9E4 CDR 2 Light Chain SEQ ID NO:49 9E4 CDR 3 Light Chain SEQ ID NO:50 GM63 Epitope 126-138 SEQ ID NO:51 GM63CDR 1 Heavy Chain SEQ ID NO:52 GM63 CDR 2 Heavy Chain SEQ ID NO:53 GM63 CDR 3 Heavy Chain SEQ ID NO:54 GM63 CDR 1 Light Chain SEQ ID NO:55 GM63CDR 2 Light Chain SEQ ID NO:56 GM63CDR 3 Light Chain SEQ ID NO:57 GM63 Heavy Chain Variable Domain SEQ ID NO:58 GM63 Light Chain Variable Domain SEQ ID NO:59 GM63 IgG1 Heavy Chain Constant domain SEQ ID NO:60 GM63 Kappa Light Chain Constant domain SEQ ID NO:61 2E6 Epitope 126-140 SEQ ID NO:62 2E6 CDR 1 Heavy Chain SEQ ID NO:63 2E6 CDR 2 Heavy Chain SEQ ID NO:64 2E6 CDR 3 Heavy Chain SEQ ID NO:65 2E6 CDR 1 Light Chain SEQ ID NO:66 2E6 CDR 2 Light Chain SEQ ID NO:67 2E6 CDR 3 Light Chain SEQ ID NO:68 2E6 Heavy Chain Variable Domain SEQ ID NO:69 2E6 Light Chain Variable Domain SEQ ID NO:70 ch2E6 Heavy Chain Variable Domain SEQ ID NO:71 ch2E6 Light Chain Variable Domain SEQ ID NO:72 2E6 HLD 1 Heavy Chain Variable Domain SEQ ID NO:73 2E6 HLD 1Light Chain Variable Domain SEQ ID NO:74 2E6 HLD 2 Heavy Chain Variable Domain SEQ ID NO:75 2E6 HLD 2 Light Chain Variable Domain SEQ ID NO:76 2E6 HLD 3 Heavy Chain Variable Domain SEQ ID NO:77 2E6 HLD 3 Light Chain Variable Domain SEQ ID NO: 78 D 1.2 CDR 1 Light Chain SEQ ID NO:79 D 1.2CDR 2 Light Chain SEQ ID NO:80 D 1.2CDR 3 Light Chain SEQ ID NO:81 D 1.2CDR 1 Heavy Chain SEQ ID NO:82 D 1.2 CDR 2 Heavy Chain SEQ ID NO:83 D 1.2 CDR 3 Heavy Chain SEQ ID NO:84 D 1.2 Light Chain Variable Domain SEQ ID NO:85 D 1.2 Heavy Chain Variable Domain SEQ ID NO:86 C 10.2 CDR 1 Light Chain SEQ ID NO:87 C 10.2CDR 2 Light Chain SEQ ID NO:88 C 10.2CDR 3 Light Chain SEQ ID NO:89 C 10.2CDR 1 Heavy Chain SEQ ID NO:90 C 10.2 CDR 2 Heavy Chain SEQ ID NO:91 C 10.2 CDR 3 Heavy Chain

SEQ ID NO:92 C 10.2 Light Chain Variable Domain SEQ ID NO:93 C 10.2 Heavy Chain Variable Domain SEQ ID NO:94 CDR1 VL 7C4 SEQ ID NO:95 CDR1 VL 7A10 &8D9 SEQ ID NO:96 CDR3 VL L3 SEQ ID NO:97 CDR1 VH 7C4 SEQ ID NO:98 CDR2 VH 5A1 SEQ ID NO:99 CDR2 VH 9G11 SEQ ID NO:100 CDR2 VH 9C12 SEQ ID NO:101 CDR3 VH 5A1 SEQ ID NO:102 CDR3 VH 9D7 SEQ ID NO:103 CDR3 VH 7A10 &8D9 SEQ ID NO:104 Full length VL 5A1 SEQ ID NO:105 Full length VH 5A1 SEQ ID NO:106 Full length VL 9D7 SEQ ID NO:107 Full length VH 9D7 SEQ ID NO:108 Full length VL 9G11 SEQ ID NO:109 Full length VH 9G11 SEQ ID NO:110| Full length VL 7C4 SEQ ID NO:111| Full length VH 7C4 SEQ ID NO:112 Full length VL L3 SEQ ID NO:113 Full length VH L3 SEQ ID NO:114 Full length VL 7A10 SEQ ID NO:115 Full length VH 7A10 SEQ ID NO:116 Full length VL 8D9 SEQ ID NO:117 Full length VH 8D9 SEQ ID NO:118 Full length VL 9C12 SEQ ID NO:119 Full length VH 9C12 SEQ ID NO:120 Full length VL 6B6 SEQ ID NO:121 Full length VH 6B6

5

10

Further Embodiments of the Invention 1. An alpha-synuclein binding monoclonal antibody, or an antigen-binding fragment thereof, for use in inhibiting aggregation of tau. 2. The monoclonal antibody, or antigen-binding fragment thereof, according 5 to item 1, wherein the antibody binds aggregated soluble forms of alpha synuclein 3. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1 or 2, wherein inhibition of tau aggregation is in vivo or in vitro. 4. The monoclonal antibody, or antigen-binding fragment thereof, according 10 to any one of the preceding items, wherein said alpha-synuclein antibody is administered to a patient with Alzheimer's disease. 5. The monoclonal antibody, or antigen-binding fragment thereof, according to any one of the preceding items, wherein the patient does not have Lewy body variant of Alzheimer's disease or combined Parkinson and Alz 15 heimer's disease. 6. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-3 wherein said alpha-synuclein antibody is administered to a patient with a taupahty selected from the group comprising Argyrophilic Grain Disease (AGD), Psychosis, particularly Psychosis due to AD or 20 Psychosis in patients with AD, psychiatric symptoms of patients with Lewy body dementia, Progressive Supranuclear Palsy (PSP), Frontotemporal dementia (FTD or variants thereof), TBI (traumatic brain injury, acute or chronic), Corticobasal Degeneration (CBD), Picks Disease, Primary age related tauopathy (PART), Neurofibrillary tangle-predominant senile de 25 mentia, Dementia pugilistica, Chronic traumatic encephalopathy, stroke, stroke recovery, neurodegeneration in relation to Parkinson's disease, Parkinsonism linked to chromosome, Lytico-Bodig disease (Parkinson-de mentia complex of Guam), Ganglioglioma and gangliocytoma, Meningio angiomatosis, Postencephalitic parkinsonism, Subacute sclerosing 30 panencephalitis, Huntington's disease, lead encephalopathy, tuberous sclerosis, Hallervorden-Spatz disease and lipofuscinosis. More typically, the taupathy is selected from the group consisting of Alzheimer's disease, Argyrophilic Grain Disease (AGD), Psychosis, particularly Psychosis due to AD or Psychosis in patients with AD, psychiatric symptoms of patients with Lewy body dementia, Progressive Supranuclear Palsy (PSP), Fronto temporal dementia (FTD or variants thereof), TBI (traumatic brain injury, acute or chronic), Corticobasal Degeneration (CBD), and Picks Disease. 5 7. The monoclonal antibody, or antigen-binding fragment thereof, according to any one of the preceding items, wherein said alpha-synuclein antibody binds to the C-terminal part of alpha-synuclein 8. The monoclonal antibody, or antigen-binding fragment thereof, according to item 7, wherein the antibody binds to an epitope within the C-terminal 10 amino acids 110-140 of human alpha-synuclein.

9. The monoclonal antibody, or antigen-binding fragment thereof, according to items 7 or 8, wherein said epitope is within amino acids 112-117, 112 115, 118-126, 126-138 or 136-140 of human alpha-synuclein (SEQ ID NO 10).

15 10. The monoclonal antibody according to any one of the preceding items, wherein said antibody binds an epitope within amino acids 112-117 (SEQ ID NO:9 (ILEDMP)) of human alpha-synuclein (SEQ ID NO:10), or antigen-binding fragment thereof that binds said epitope.

11. The monoclonal antibody, or antigen-binding fragment thereof, according 20 to according any one of the preceding items, wherein said antibody is capable of competing with an antibody comprising the light chain variable domain of SEQ ID NO:8 and the heavy chain variable domain of SEQ ID NO:7, 30, 31 or 32 for binding to said epitope.

12. The monoclonal antibody according to any one of the preceding items, 25 wherein said antibody is capable of specifically binding to an epitope within amino acids 112-115 (SEQ ID NO:19 (ILED) of human alpha synuclein (SEQ ID NO:10)), or antigen-binding fragment thereof that binds said epitope.

13. The monoclonal antibody, or antigen-binding fragment thereof, according to any one of the preceding items, wherein said antibody is capable of competing with an antibody comprising the heavy chain variable domain of SEQ ID NO:26 and the light chain variable domain of SEQ ID NO:27 5 for binding to said epitope

14. The monoclonal antibody according to item 9, wherein said antibody binds an epitope within amino acids 118-126 (such as SEQ ID NO:36 (NEAYE)) of human alpha-synuclein (SEQ ID NO:10), or antigen-binding fragment thereof that binds said epitope.

10 15. The monoclonal antibody, or antigen-binding fragment thereof, according to item 9, wherein said antibody is capable of competing with an antibody comprising the heavy chain variable domain of SEQ ID NO:42 and the light chain variable domain of SEQ ID NO:43 for binding to said epitope.

16. The monoclonal antibody according to item 9, wherein said antibody binds 15 an epitope within amino acids 126-138 (SEQ ID NO:50 (EMPSEEGYQD YEP) of human alpha-synuclein (SEQ ID NO:10), or antigen-binding fragment thereof that binds said epitope.

17. The monoclonal antibody, or antigen-binding fragment thereof, according to item 9, wherein said antibody is capable of competing with an antibody 20 comprising the heavy chain variable domain of SEQ ID NO:57 and the light chain variable domain of SEQ ID NO:58 for binding to said epitope.

18. The monoclonal antibody according to item 9, wherein said antibody binds an epitope within amino acids 126-140 (SEQ ID NO:61 (EMPSEEGYQD YEPEA) of human alpha-synuclein (SEQ ID NO:10), or antigen-binding 25 fragment thereof that binds said epitope.

19. The monoclonal antibody, or antigen-binding fragment thereof, according to item 18, wherein said antibody is capable of competing with an antibody comprising the heavy chain variable domain of SEQ ID NO:68 and the light chain variable domain of SEQ ID NO:69 for binding to said epitope.

20. The monoclonal antibody according to any one of the preceding items comprising or consisting of an intact antibody.

5 21. The monoclonal antibody according to any one of the preceding items wherein the monoclonal antibody is selected from the group consisting of antibodies of subtype IgG1, IgG2, IgG3 or IgG4.

22. The monoclonal antibody, or antigen-binding fragment thereof, according to any one of the preceding items comprising or consisting of an antigen 10 binding fragment selected from the group consisting of Fv fragments (e.g. single chain Fv and disulphide-bonded Fv), Fab-like fragments (e.g. Fab fragments, Fab'fragments and F(ab)2 fragments) and domain antibodies (e.g. single VH variable domains or VL variable domains).

23. The monoclonal antibody, or antigen-binding fragment thereof, according 15 to any one of the preceding items wherein the antibody or antigen-binding fragment exhibits one or more of the following properties: a) a binding affinity (KD) for alpha-synuclein of between 0.5-10 nM, such as 1-5 nM or 1-2 nM; b) capability of inhibiting protease truncation of alpha-synuclein fibrils; 20 c) capability of reversing impairment in basal synaptic transmission in F28-snca transgenic mice; d) capability of reducing levels of alpha-synuclein in the mouse hippocampus as measured by in vivo microdialysis; e) capability, when administered chronically, to restore motor function in 25 a rat model of Parkinson's disease; f) Capability to prevent seeding of alpha-synuclein (such as accumula tion of insoluble phosphorylated alphasynuclein in vitro and/or in a mouse model of Parkinson's disease); and/or g) Capability to bind truncated alpha-synuclein in a human brain.

24. The monoclonal antibody, or antigen-binding fragment thereof, according to any one of the previous items that is a human, humanized, recombinant or chimeric antibody.

25. The monoclonal antibody, or antigen-binding fragment thereof, according 5 to items 1-11 wherein the antibody, or a fragment thereof, comprises: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:2; 10 (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID 15 NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6.

26. The monoclonal antibody, or antigen-binding fragment thereof, according to item 25, comprising the heavy chain variable domain of SEQ ID NO:7 20 or the light chain variable domain of SEQ ID NO:8.

27. The monoclonal antibody, or antigen-binding fragment thereof, according to item 25, comprising a heavy chain consisting of a variable domain of SEQ ID NO:7 and a light chain consisting of a variable domain of SEQ ID NO:8.

25 28. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-11 wherein the antibody, or a fragment thereof, comprises: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID 30 NO:33;

(c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; 5 (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6.

29. The monoclonal antibody, or antigen-binding fragment thereof, according 10 to item 28, comprising the heavy chain variable domain of SEQ ID NO:30 or the light chain variable domain of SEQ ID NO:8.

30. The monoclonal antibody, or antigen-binding fragment thereof, according to item 28, comprising a heavy chain consisting of a variable domain of SEQ ID NO:30 and a light chain consisting of a variable domain of SEQ 15 ID NO:8.

31. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-11 wherein the antibody, or a fragment thereof, comprises: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; 20 (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:34; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID 25 NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6.

32. The monoclonal antibody, or antigen-binding fragment thereof, according to item 31, comprising the heavy chain variable domain of SEQ ID NO:31 or the light chain variable domain of SEQ ID NO:8.

33. The monoclonal antibody, or antigen-binding fragment thereof, according 5 to item 31, comprising a heavy chain consisting of a variable domain of SEQ ID NO:31 and a variable domain of SEQ ID NO:8.

34. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-11 wherein the antibody, or a fragment thereof, comprises: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID 10 NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:35; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; 15 (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID 20 NO:6.

35. The monoclonal antibody, or antigen-binding fragment thereof, according to item 34, comprising the heavy chain variable domain of SEQ ID NO:32 or the light chain variable domain of SEQ ID NO:8.

36. The monoclonal antibody, or antigen-binding fragment thereof, according 25 to item 34, comprising a heavy chain consisting of a variable domain of SEQ ID NO:32 and a variable domain of SEQ ID NO:8.

37. The monoclonal antibody, or antigen-binding fragment thereof, according to any items 1-13 wherein the antibody, or a fragment thereof, comprises:

(a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:20; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:21; 5 (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:22; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:23; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID 10 NO:24; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:25.

38. The monoclonal antibody, or antigen-binding fragment thereof, according to item 37, comprising the heavy chain variable domain of SEQ ID NO:26 15 or the light chain variable domain of SEQ ID NO:27.

39. The monoclonal antibody, or antigen-binding fragment thereof, according to item 37, comprising a heavy chain consisting of a variable domain of SEQ ID NO:26 and a variable domain of SEQ ID NO:27.

20 40. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-9 and items 16-17, wherein said alpha-synuclein antibody com prises: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:51; 25 (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:52; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:53; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID 30 NO:54;

(e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:55; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:56. 5 41. The monoclonal antibody, or antigen-binding fragment thereof, according to item 40, comprising the heavy chain variable region of SEQ ID NO:57 or the light chain variable region of SEQ ID NO:58. 42. The monoclonal antibody, or antigen-binding fragment thereof, according to item 40, comprising the heavy chain variable region of SEQ ID NO:57 10 and the light chain variable region of SEQ ID NO:58. 43. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-9 and items 14-15, wherein said alpha-synuclein antibody com prises: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID 15 NO:44; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:45; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:46; 20 (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:47; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:48; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID 25 NO:49. 44. The monoclonal antibody, or antigen-binding fragment thereof, according to item 43, comprising the heavy chain variable region of SEQ ID NO:42 or the light chain variable region of SEQ ID NO:43. 45. The monoclonal antibody, or antigen-binding fragment thereof, according 30 to item 43, comprising the heavy chain variable region of SEQ ID NO:42 and the light chain variable region of SEQ ID NO:43.

46. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-9 and items 18-19, wherein said alpha-synuclein antibody com prises: o SEQ ID NO: 94 or an amino acid sequence having no more than 5 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; o SEQ ID NO: 66 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid 10 differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and o SEQ ID NO: 67 or an amino acid sequence having with no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no 15 more than 1 amino acid difference. 47. The monoclonal antibody, or antigen-binding fragment thereof, according to Item 46 comprising a light chain variable region comprising the CDRs of SEQ ID NOs 94,66 and 67. 48. The monoclonal antibody, or antigen-binding fragment thereof, according 20 to items 46 or 47, wherein said alpha-synuclein antibody comprises: o SEQ ID NO: 97 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; 25 o SEQ ID NO: 63 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and * SEQ ID NO: 64 or an amino acid sequence having no more 30 than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference.

49. The monoclonal antibody, or antigen-binding fragment thereof, according to Item 48 comprising a heavy chain variable region comprising the CDRs of SEQ ID NOs 97, 63 and 64. 50. The monoclonal antibody, or antigen-binding fragment thereof, according 5 to Items 46 or 47 comprising a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 110. 51. The monoclonal antibody, or antigen-binding fragment thereof, according to Items 48 or 49 comprising a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:111. 10 52. The monoclonal antibody, or antigen-binding fragment thereof, according items 50 and 51comprising a light chain variable region comprising or con sisting of the amino acid sequence of SEQ ID NO:110 and heavy a chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:111. 15 53. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-9 and items 18-19, wherein said alpha-synuclein antibody com prises: o SEQ ID NO: 95 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ 20 ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; o SEQ ID NO: 66 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no 25 more than 1 amino acid difference; and o SEQ ID NO: 67 or an amino acid sequence having with no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference. 30 54. The monoclonal antibody, or antigen-binding fragment thereof, according to Item 53 comprising a light chain variable region comprising the CDRs of SEQ ID NOs 95,66 and 67.

55. The monoclonal antibody, or antigen-binding fragment thereof, according the preceding items 53 or 54 comprising a heavy chain variable region com prising the following CDRs: o SEQ ID NO: 62 or an amino acid sequence having no more than 5 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; o SEQ ID NO: 63 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ 10 ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and o SEQ ID NO: 103 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid dif 15 ferences, or no more than 1 amino acid difference. 56. The monoclonal antibody, or antigen-binding fragment thereof, according to Item 55 comprising a heavy chain variable region comprising the CDRs of SEQ ID NOs 62, 63 and 103. 57. The monoclonal antibody, or antigen-binding fragment thereof, according 20 to Items 53 or 54 comprising a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 114. 58. The monoclonal antibody, or antigen-binding fragment thereof, according to Items 55 or 56 comprising a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:115. 25 59. The monoclonal antibody, or antigen-binding fragment thereof, according items 57 and 58 comprising a light chain variable region comprising or con sisting of the amino acid sequence of SEQ ID NO:114 and heavy a chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:115. 30 60. The monoclonal antibody, or antigen-binding fragment thereof, according to Items 53 or 54 comprising a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 116.

61. The monoclonal antibody, or antigen-binding fragment thereof, according to Items 55 and 56 comprising a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:117. 62. The monoclonal antibody, or antigen-binding fragment thereof, according 5 items 60 and 61 comprising a light chain variable region comprising or con sisting of the amino acid sequence of SEQ ID NO:116 and heavy a chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:117. 63. The monoclonal antibody, or antigen-binding fragment thereof, according 10 to items 1-9 and items 18-19, wherein said alpha-synuclein antibody com prises: o SEQ ID NO: 65 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more 15 than 1 amino acid difference; o SEQ ID NO: 66 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and 20 o SEQ ID NO: 96 or an amino acid sequence having with no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference. 64. The monoclonal antibody, or antigen-binding fragment thereof, according 25 to Item 63 comprising a light chain variable region comprising the CDRs of SEQ ID NOs 65, 66 and 96. 65. The monoclonal antibody, or antigen-binding fragment thereof, according the preceding items 63 or 64 comprising a heavy chain variable region com prising the following CDRs: 30 o SEQ ID NO: 62 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; o SEQ ID NO: 63 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and 5 o SEQ ID NO: 64 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference. 66. The monoclonal antibody, or antigen-binding fragment thereof, according 10 to Item 65 comprising a heavy chain variable region comprising the CDRs of SEQ ID NOs 62, 63 and 64. 67. The monoclonal antibody, or antigen-binding fragment thereof, according to Items 63 or 64 comprising a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 112. 15 68. The monoclonal antibody, or antigen-binding fragment thereof, according to Items 65 or 66 comprising a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:113. 69. The monoclonal antibody, or antigen-binding fragment thereof, according items 67 and 68 comprising a light chain variable region comprising or con 20 sisting of the amino acid sequence of SEQ ID NO:112 and heavy a chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:113. 70. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-9 and items 18-19, wherein said alpha-synuclein antibody com 25 prises: o SEQ ID NO: 65 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; 30 o SEQ ID NO: 66 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and o SEQ ID NO: 67 or an amino acid sequence having with no more than 4 amino acid differences, or no more than 3 amino acid 5 differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference. 71. The monoclonal antibody, or antigen-binding fragment thereof, according to Item 70 comprising a light chain variable region comprising the CDRs of SEQ ID NOs 65,66 and 67. 10 72. use according the preceding items 70 or 71 comprising a heavy chain var iable region comprising the following CDRs: o SEQ ID NO: 62 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more 15 than 1 amino acid difference; o SEQ ID NO: 98 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and 20 o SEQ ID NO: 101 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid dif ferences, or no more than 1 amino acid difference. 73. The useaccording to Item 72 comprising a heavy chain variable region com 25 prising the CDRs of SEQ ID NOs 62, 98 and 101 74. The monoclonal antibody, or antigen-binding fragment thereof, according to Items 70 or 71 comprising a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 104. 75. The use Items 72 or 73 comprising a heavy chain variable region compris 30 ing or consisting of the amino acid sequence of SEQ ID NO:105.

76. The monoclonal antibody, or antigen-binding fragment thereof, according items 74 and 75 comprising a light chain variable region comprising or con sisting of the amino acid sequence of SEQ ID NO:104 and heavy a chain variable region comprising or consisting of the amino acid sequence of 5 SEQ ID NO:105. 77. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-9 and items 18-19, wherein said alpha-synuclein antibody com prises: o SEQ ID NO: 65 or an amino acid sequence having no more than 10 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; o SEQ ID NO: 66 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid 15 differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and o SEQ ID NO: 67 or an amino acid sequence having with no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no 20 more than 1 amino acid difference. 78. The monoclonal antibody, or antigen-binding fragment thereof, according to Item 77 comprising a light chain variable region comprising the CDRs of SEQ ID NOs 65,66 and 67. 79. The monoclonal antibody, or antigen-binding fragment thereof, according 25 the preceding items 77 or 78 comprising a heavy chain variable region com prising the following CDRs: o SEQ ID NO: 62 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more 30 than 1 amino acid difference; o SEQ ID NO: 99 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and 5 o SEQ ID NO: 64 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference. 80. The monoclonal antibody, or antigen-binding fragment thereof, according 10 to Item 79 comprising a heavy chain variable region comprising the CDRs of SEQ ID NOs 62,99 and 64. 81. The monoclonal antibody, or antigen-binding fragment thereof, according to Items 77 or 78 comprising a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 108. 15 82. The monoclonal antibody, or antigen-binding fragment thereof, according to Items 79 or 80 comprising a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:109. 83. The monoclonal antibody, or antigen-binding fragment thereof, according items 81 and 82 comprising a light chain variable region comprising or con 20 sisting of the amino acid sequence of SEQ ID NO:108 and heavy a chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:109. 84. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-9 and items 18-19, wherein said alpha-synuclein antibody com 25 prises: o SEQ ID NO: 65 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; 30 o SEQ ID NO: 66 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and o SEQ ID NO: 67 or an amino acid sequence having with no more than 4 amino acid differences, or no more than 3 amino acid 5 differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference. 85. The monoclonal antibody, or antigen-binding fragment thereof, according to Item 84 comprising a light chain variable region comprising the CDRs of SEQ ID NOs 65,66 and 67. 10 86. The monoclonal antibody, or antigen-binding fragment thereof, according the preceding items 84 or 85 comprising a heavy chain variable region com prising the following CDRs: o SEQ ID NO: 62 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ 15 ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; o SEQ ID NO: 100 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no 20 more than 1 amino acid difference; and o SEQ ID NO: 64 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference. 25 87. The monoclonal antibody, or antigen-binding fragment thereof, according to Item 86 comprising a heavy chain variable region comprising the CDRs of SEQ ID NOs 62, 100 and 64. 88. The useaccording to Items 84 or 85 comprising a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 118. 30 89. The monoclonal antibody, or antigen-binding fragment thereof, according to Items 86 or 87 comprising a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:119.

90. The monoclonal antibody, or antigen-binding fragment thereof, according to any one of the preceding items 88 and 89 comprising a light chain vari able region comprising or consisting of the amino acid sequence of SEQ ID NO:118 and heavy a chain variable region comprising or consisting of the 5 amino acid sequence of SEQ ID NO:119. 91. The monoclonal antibody, or antigen-binding fragment thereof, according to items 1-9 and items 18-19, wherein said alpha-synuclein antibody com prises: o SEQ ID NO: 65 or an amino acid sequence having no more than 10 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; o SEQ ID NO: 66 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid 15 differences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and o SEQ ID NO: 67 or an amino acid sequence having with no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid differences, or no 20 more than 1 amino acid difference. 92. The monoclonal antibody, or antigen-binding fragment thereof, according to Item 91 comprising a light chain variable region comprising the CDRs of SEQ ID NOs 65,66 and 67. 93. The monoclonal antibody, or antigen-binding fragment thereof, according 25 the preceding items 91 or 92 comprising a heavy chain variable region com prising the following CDRs: o SEQ ID NO: 62 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more 30 than 1 amino acid difference; o SEQ ID NO: 63 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differ ences, or no more than 2 amino acid differences, or no more than 1 amino acid difference; and 5 o SEQ ID NO: 102 or an amino acid sequence having no more than 4 amino acid differences, or no more than 3 amino acid differences, or no more than 2 amino acid dif ferences, or no more than 1 amino acid difference. 94. The monoclonal antibody, or antigen-binding fragment thereof, according 10 to Item 93 comprising a heavy chain variable region comprising the CDRs of SEQ ID NOs 62, 63 and 102 95. The monoclonal antibody, or antigen-binding fragment thereof, according to Items 91 or 92 comprising a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 106. 15 96. The antibody or antigen-binding fragment thereof according to Items 93 or 94 comprising a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:107. 97. The monoclonal antibody, or antigen-binding fragment thereof, according items 95 or 96 comprising a light chain variable region comprising or con 20 sisting of the amino acid sequence of SEQ ID NO:106 and heavy a chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:107. 98. A nucleic acid encoding the antibody or the fragment according to any one of items 25-97. 25 99. A pharmaceutical composition comprising the monoclonal antibody, or an antigen-binding fragment thereof, according to any one of the previous items or the preparation of any one of items 25-97, and a pharmaceutical acceptable carrier. 100. A method of treating a disease according to items 5 or 6 in a sub 30 ject, said method comprising administering the monoclonal antibody or antigen-binding fragment thereof of any of items 1-97 to said subject in an effective amount.

101. The method according to item 100, wherein the treatment is chronic 102. The method according to item 101, wherein the chronic treatment is for at least 2 weeks. 5 103. The method according to item 100, wherein the subject is human. 104. A kit comprising the antibody, or antigen-binding fragment thereof, according to items 1-97 for use in a method according to item 100. 105. The monoclonal antibody, or antigen-binding fragment thereof, ac cording to items 5 or 6, wherein the monoclonal antibody of items 1-97 is 10 detectably labelled. 106. The monoclonal antibody, or antigen-binding fragment thereof, ac cording to of item 105, wherein said detectable label is a fluorescent label, a chemoluminescent label, a paramagnetic label, a radioisotopic label or an enzyme label. 15 107. The monoclonal antibody, or antigen-binding fragment thereof, ac cording to of items 105-106 for use in detecting or measuring the pres ence or amount of said alpha-synuclein in the brain or any other organ or body fluid of a subject. 108. The monoclonal antibody, or antigen-binding fragment thereof, ac 20 cording to items 105-107, wherein said detection or measurement com prises in vivo imaging of said anti-synulclein antibody bound to said alpha synuclein. 109. The monoclonal antibody, or antigen-binding fragment thereof, ac cording to items 105-108, wherein said detection or measurement com 25 prises ex vivo imaging of said anti-synuclein antibody or said antigen binding fragment thereof, bound to said alpha-synuclein. 110. Use of a monoclonal antibody, or antigen-binding fragment thereof, according to any one of items 1-97 for use in the manufacturing of a medicament for treating, diagnosing or imaging a disease according to 30 items 5 or 6. 111. A method of delaying the progression of a disease according to items 5 or 6 in a patient, said method comprising reducing or attenuating the accumulation of pathological tau protein in said patient by administer ing an antibody as defined in items 1-97

EXAMPLES

Example 1: Antibody Screening

1. Immunogen and ligand production

[000135] The following proteins were acquired or produced for use as 5 immunogens shown in Fig 1. The mice were immunized with three immunogens: full length recombinant human alpha-synuclein fibrils; human alpha-synuclein recombinant protein containing amino acids 1-60 (Rpeptide, Bogart, Georgia) and human alpha-synuclein recombinant protein containing amino acids 1-119. To make the fibrils from the full length the alpha-synuclein alyophilized product from 10 Rpeptide, Bogart, Georgia (Catalog number S-1001-2) was used. This was dissolved in 20 mM tris and 300 mM NaCl buffer at concentration of 1 mg/ml protein. To make the fibrils the protein solution was incubated 170 pl aliquots in 96 well plate with a 70 pm diameter ceramic bead in each well at 200 rpm in Vortemp 56 shaker incubator (Labnet International, Edison, NJ, USA), at 370C 15 for 7 days, and the formation of fibrils was followed by adding thioflavin T and measuring fluorescence in one of the wells. The recombinant alpha-synuclein containing amino acids 1-60 was dissolved in water to give a concentration of 1 mg/mi.

[000136] The recombinant alpha-synuclein containing amino acids 1-119 20 was made using the following construct: A synthetic gene coding for a 6 amino acid Histidine tag, followed by factor Xa cleavage site and sequence coding for human alpha-synuclein amino acids 1-119: MAHHHHHHIE GRMDVFMKGL SKAKEGVVAA AEKTKQGVAE AAGKTKEGVL YVGSKTKEGV VHGVATVAEK TKEQVTNVGG AVVTGVTAVA QKTVEGAGSI 25 AAATGFVKKD QLGKNEEGAP QEGILEDMPV D (SEQ ID NO:16) was synthezised by Genscript and cloned into Ndel-Xhol site in pET24a(+) expression vector (Novagen).

[000137] The expression vector was transformed into E. coli BL21 and a single colony picked for expression using the overnight express autoinduction 30 system from Novagen (User protocol TB383 rev. H1005). The scale was 500 ml of final culture volume. Cells were harvested by centrifugation 10 min at 6000g and subsequently lyzed using BugBuster protein extraction Reagent (User protocol TB245 Rev. 0304). After lysis the sample was cleared by centrifugation and the supernatant used for further purification.

[000138] The His-tagged protein was purified on a 5ml HisTrap column (GE 5 healthcare) equilibrated in 20 mM Sodium phosphate pH7.5, 1 M NaCl (A-buffer). After sample application and wash using A-buffer the protein was eluted in a gradient to 0.25 M Imidazole in A-buffer over 20 column volumes. Fractions of 5 ml were collected and analyzed by SDS-PAGE. Fractions with the protein of interest was pooled, concentrated and applied to an S200 (26/60) size exclusion 10 column (GE healthcare) in 10 mM tris pH 7.4, 300 mM NaCl. Again fractions were pooled according to presence in SDS-PAGE of a band with expected size.

[000139] To remove the N-terminal tag, the purified his-tagged alpha synuclein 1-119 was incubated with factor Xa in a 1:50 ration using the Novagen kit (69037-3FRX). After overnight incubation, the factor Xa was removed 15 batchwise using Xarrest agarose. The cleaved alpha-synuclein 1-119 was finally purified by permissive HisTrap chromatography as described above. From the flow through the purified alpha-synuclein 1-119 was obtained and concentrated to -400 pg/ml using centricon concentration devises.

[000140] Alpha-synuclein (Rpeptide) was rehydrated in PBS at 2 mg/ml and 20 peroxynitirite (1OOpL/mg protein) was added dropwise while mixing. The nitrosylated alpha-synuclein was then dialyzed in 5L PBS and stored at -20°C.

[000141] Dopamine was used to oxidize alpha-synuclein. Equal volumes of a 200uM solution of Dopamine-HCL (Sigma P5244) prepared in 10mM PBS, pH7.4 and a 28pM solution of alpha-synuclein (Rpeptide) in 10mM PBS, pH7.4 25 were combined. The resulting 14 uM alpha-synuclein/100 uM Dopamine were incubated at 370C O/N (over night). The oxidized alpha-synuclein was then dialyzed in PBS and stored at -20°C.

[000142] Different native and chimeric versions of synuclein proteins were produced in order to screen a diverse library of anti-alpha-synuclein antibodies.

Screening constructs included the following: human, mouse, rat and cynomolgus monkey alpha-synuclein, human Beta-synuclein, Human Gamma-synuclein and lastly an alpha-synuclein derivative that lacked residues 120-140 of alpha synuclein. In addition, a series of 4 shuffle constructs: A-Syn-AAKK-BAP, A-Syn 5 BAAK-BAP, A-Syn-BBAA-BAP, A-Syn-BBKK-BAP (SEQ ID Nos:11-14) were produced. These constructs contained linear stretches of human alpha-synuclein (A), human Beta-synuclein (B) and chicken alpha-synuclein (K). Gene were cloned containing a Biotin Acceptor Peptide (BAP) tag fused to the C-terminus of the ligands in order to facilitate site specific biotinylation of each of the ligands. 10 The bioytinylation allowed for attachment of the ligands to beads used in the soluble ELISA format. Mammalian expression vectors were constructed carrying the different alpha-synuclein BAP tag fusion constructs (ASynBAP). The ligands were expressed in HEK 293 cells using transient transfection (Genmab A/S).

2. Immunization

15 [000143] Antibodies HuMab-Synuclein were derived from the immunizations of HuMAb mouse strain HCol7-BALB/c and HCol2-BALB/c mice, double knock out for the mouse immunoglobulin (Ig) heavy and mouse kappa light chain, which prevents the expression of antibodies that are completely murine (human monoclonal antibody; Medarex Inc., San Jose, CA, USA). The various mouse 20 strains were made transgenic by the insertion of human Ig heavy and human Ig kappa light chain loci and differ in the number of human VH (variable domain of heavy chain) and VL (variable domain of light chain) genes.

[000144] 48 Mice were immunized alternating intraperitoneally (IP) with 20 pg antigens and subcutaneously (SC, at the tailbase) with the same immunogen, 25 with an interval of 14 days. A maximum of eight immunizations were performed, 4 IP and 4 SC.

[000145] The first immunization was performed with alpha-synuclein immunogens in complete Freund's adjuvant (CFA; Difco Laboratories, Detroit, MI, USA), the following immunizations in incomplete Freund's adjuvant (IFA). When 30 serum titers were found to be sufficient (dilution of serum of 1/50 or lower found positive in antigen specific screening assay as described in herein above on at least two sequential, biweekly, screening events), mice were additionally boosted twice intravenously (IV) with 10 pg alpha-synuclein immunogen protein in 100 pL PBS, four and three days before fusion.

5 [000146] The immunization protocols are shown in Fig 1.

[000147] Antibody GM37 came from an immunization protocol where human full length a-Synuclein-fibrils was used, alternating with alpha-synuclein C- terminally truncated forms with amino acids 1-60 and 1-119.

[000148] Antibody GM285 came from an immunization protocol where 10 Human a-Synuclein-monomer 1-140 was used for the first 4 immunizations. If there was no titer, the immunization was continued with fibrils (ip/sc), otherwise it was continued with monomer.

3. HuMab hybridoma generation

[000149] HuMAb mice with sufficient antigen-specific titer development as 15 defined above were sacrificed and the spleen and lymph nodes flanking the abdominal aorta and caval vein were collected. Fusion of splenocytes and lymph node cells with a mouse myeloma cell line was done by electrofusion using a CEEF 50 Electrofusion System (Cyto Pulse Sciences, Glen Burnie, MD, USA), essentially according to the manufacturer's instructions. Fused cells were seeded 20 in fusion medium containing 10% Fetal Clone I Bovine serum (Perbio), 1 mM sodium pyruvate (Cambrex), 0.5 U/mL penicillin, 0.5 U/mL streptomycin (Cambrex), 50 pM 2-mercaptoethanol (Invitrogen), 600 ng/mL interleukin 6 (IL-6) (Strathmann), 1 x HAT (Sigma) and 0.5 mg/mL kanamycin (Invitrogen) in HyQ mADCF-Mab (Perbio). After ten days, supernatant was harvested and cells were 25 refreshed with harvest medium, containing 10 % Fetal Clone I Bovine serum, 0.5 U/mL penicillin, 0.5 U/mL streptomycin, 600 ng/mL IL-6 and 1 x proHT (Cambrex) in HyQ mADCF-Mab. Supernatants of the hybridoma cultures were screened by primary screening assays. Supernatants were characterized for binding to eight different ligands. These included 4 orthologs: human, mouse, rat and 30 cynomologus monkey, human alpha-synuclein Beta-synuclein and human

Gamma-synuclein (SEQ ID NOs 37-41) and lastly they were tested for their ability to bind to a human alpha-synuclein derivative that lacked residues 120-140 of alpha-synuclein.

[000150] The screening of anti-alpha-synuclein antibodies was performed 5 using a high throughput suspension ELISA format using automated liquid handling systems (Genmab A/S). The reading of the plates was performed by two systems, the FMAT 8200 from Applied Biosystems was used to read 384 well plates and the ImageXpress Velos Cytometer from Molecular Devices was used to read the 1536 well plates.

10 [000151] In the primary screen clones were characterized by their ability to bind 8 different ligands. These included a series of 4 shuffle constructs: A-Syn AAKK-BAP, A-Syn-BAAK-BAP, A-Syn-BBAA-BAP, A-Syn-BBKK-BAP (SEQ ID NOs:11-14), alpha-synuclein 120-140 deletion-BAP, nitrated human alpha synuclein-BAP and oxidized human alpha-synuclein-BAP.

15 [000152] In short, the sera or supernatant potentially containing alpha synuclein specific antibodies were added to the beads to allow binding to alpha Synuclein and/or alpha-synuclein derived constructs. The binding of the anti alpha-synuclein antibodies is detected using a fluorescent conjugate, DyLight649 conjugated goat antihuman IgG, Fc specific. Two known mouse anti-alpha 20 synuclein antibodies, LB509 and Syn211, were included in screenings as positive controls. To ensure specific detection of alpha-synuclein antibodies, an anti alpha-synuclein sera pool was used as a negative control in the 384 well format titer screening while human ChromPure IgG is used in the 1536 well format 8 bead based assay.

25 [000153] Hybridoma cells from the best primary wells were seeded in semisolid medium made from 40% CloneMedia (Genetix, Hampshire, UK) and 60% HyQ 2x complete medium (Hyclone, Waltham, USA). For each primary well, a well of a Genetix black 6-well plate was seeded. From each well, 25 sub clones were picked, using the ClonePix system (Genetix). The sub clones were picked in 30 harvest medium. After seven days, the supernatants of the sub clones were screened again for Synuclein-specific human IgG binding and the human IgG concentration was measured using Octet (Fortebio, Menlo Park, USA). From each primary well, the best sub clone was selected and expanded in expansion medium containing only 600 ng/mL IL-6, 0.5 U/mL penicillin, 0.5 U/mL 5 streptomycin and 1 x proHT. The sub clones were expanded from one 96-well plate well to one 24-well plate well to four 24-well plate wells to six 6-well plate wells. Clones derived by this process were designated as primary clones (PC).

[000154] Additional antibody binding studies were performed using Octet 384RED (Fortebio, Menlo Park, USA). HuMab antibody solutions of 2 pg/ml were 10 made by dilution in sample diluent (ForteBio, art. No. 18-5028). Amine reactive sensors (ForteBio, art.no. 18-0008) were used for immobilization of HuMabs. Prior to coupling to amine reactive sensors, HuMabs were diluted in MES pH 6.0 buffer (18-5027). Coupling was performed at 300C and 1000 rpm as follows: Amine reactive sensors were pre-wet in PBS and subsequently activated with 15 EDC/NHS(ForteBio. Art.no. 18-1033/18-1034) activation solution (according to manufacturer's instruction) for 300 seconds. Activated sensors were immobilized with HuMabs during 600 seconds.

[000155] The binding of GM37 and GM285 in Octet to recombinant human, cynomolgus and mouse alpha-synuclein, and lack of binding to human beta or 20 gamma-synuclein is shown in Figure 2.

4. Sequence analysis of the Synuclein-specific HuMab variable domains and cloning in expression vectors

[000156] Total RNA was prepared from 0.2 to 5x106 hybridoma cells and 5'-RACE-Complementary DNA (cDNA) was prepared from 100 ng total RNA, 25 using the SMART RACE cDNA Amplification kit (Clontech). VH and VL coding regions were amplified by PCR and cloned directly, in frame, in the p33G1f and p33Kappa expression vectors (containing the human IgG1/kappa constant domain encoding sequences), by ligation independent cloning (Aslanidis, C. and P.J. de Jong, Nucleic Acids Res 1990;18(20): 6069-74). For each antibody, 16 30 VL clones and 16 VH clones were sequenced. Clones with a correct Open Reading Frame (ORF) were selected for further study and expression. Vectors of all combinations of heavy chains and light chains were transiently co-expressed in FreestyleTM 293-F cells using 293fectin.

[000157] In the case of GM37 sequencing of the VH region identified an extra cysteine in the CDR3 domain at position 106. In order to eliminate the 5 possibility of misfolding and potential loss of antibody activity due to disulfide bond formation the cysteine was mutated to serine at position 106.

[000158] Comparator antibody 9E4 was generated based on the VH and VL sequence derived from hybridoma PTA-8221 (US patent 20080175838) (SEQ ID NO 42 and 43)

10 5. Expression/Purification of Antibodies

[000159] Antibodies were produced by transfection in HEK293 6E cells using the pTT5 vectors and PElpro as a transient transfection agent (National Research Council of Canada). In short, The heavy and light chains were transfected into HEK293 cells using PElpro (VWR), and cells were supplemented 15 with TN1 (Sigma) 24 hours after transfection. Cells were grown until the viability approached 50%, and yield of antibody measured by easy IgG titre (Thermo). Culture supernatant was filtered over 0.2 pm dead-end filters, loaded on 5 mL Protein A columns (rProtein A FF, Amersham Bioscience) and eluted with 0.1 M citric acid-NaOH, pH 3. The eluate was immediately neutralized with 2M Tris-HCI, 20 pH 9 and dialyzed to 12.6 mM NaH 2 PO 4, 140 mM NaCl, pH 7.4 (B.Braun), O/N. After dialysis, samples were sterile-filtered over 0.2 pm dead-end filters. Purity was determined by SDS-PAGE and concentration was measured by nephelometry and absorbance at 280 nm. Purified antibodies were aliquoted and stored at -800 C.

25 Example 2: Antibody Characterization Using Surface Plasmon Resonance

[000160] Real time binding of the antibodies to alpha-synuclein was measured using a BlAcore@ 3000. A capture surface was prepared by amine coupling a polyclonal rabbit Anti-Mouse antibody (part of Mouse Antibody Capture Kit, GE Healthcare, Cat. no: BR-1008-38) in first flow cell (Fcl) and second flow cell (Fc2) of a CM5 chip (BlAcore@). The mouse antibody was captured in Fc2 at the concentration required to achieve a ligand level of around 500RU. The baseline was allowed to stabilize for 10min before injecting analyte (ASynBAP) in Fc1-2 at 30pl/min. ASynBAP was run at 100-3200nM and 25 5 3200RU, respectively. The highest concentration in each titration series was run in duplicate. The surface was regenerated with 10mM Glycine-HCI, pH 1.7 (30sec inject) to remove captured mouse antibody and analyte in the end of each cycle. HBS-EP (GE Healthcare, Cat. No: BR-1001-88) was used as running buffer and sample diluent in all experiments and the assay was run at 250C. All 10 samples were kept at 40C before acquisition.

[000161] The response recorded in Fc1, where capture antibody had been immobilized but no Alpha-Synuclein antibody captured, was subtracted from the response in Fc2. A 1:1 or 2:1 binding algorithm was fit to the dataset using BlAevaluation software version 4.1.1. Results can be seen in Figs. 3, 4 and 5 15 showing binding of antibody GM37, GM285 and 9E4 to human alpha-synuclein.

Example 3: Epitope Mapping

[000162] Epitope mapping of the antibodies to alpha-synuclein was done with arrays of overlapping linear peptides at Pepscan (Pepscan Zuidersluisweg 2 8243 RC Lelystad, The Netherlands). The binding of antibody to each of the 20 synthesized 20 mer peptides was tested in a Pepscan based ELISA. The linear peptide array covering the entire coding sequence of alpha-synuclein, as well as all peptides with oxidized methionines or nitrosylated tyrosines, were incubated with primary antibody solution (overnight at 40C). After washing, the peptide arrays were incubated with a 1/1000 dilution of an antibody peroxidase conjugate 25 (SBA, cat. nr. 2010-05) for one hour at 250C. After washing, the peroxidase substrate 2,2'-azino-di-3-ethylbenzthiazoline sulfonate (ABTS) and 2 pl/ml of 3 percent H202 were added. After one hour, the color development was measured. The color development was quantified with a charge coupled device (CCD) camera and an image processing system. For data processing the values were 30 obtained from the CCD camera range from 0 to 3000 mAU, similar to a standard 96-well plate ELISA-reader. The results were quantified and stored into the

Peplab database. Occasionally a well contains an air-bubble resulting in a false positive value, the cards are manually inspected and any values caused by an air-bubble are scored as 0. The binding data of antibody GM37 and GM285 to peptides containing the sequence ILEDMP or ILED respectively can be seen in 5 Figure 6.

Example 4: GM37 and GM37 variants

[000163] The anti-alpha-synuclein antibodies were produced in mammalian cell culture under conditions that mimic the production conditions that will be used for producing clinical grade material for use in patients. It is well known 10 that proteins produced in this manner undergo post-translational modifications that can impact both therapeutic potency of the antibody as well as biophysical attributes that affect the stability of the antibody over time. Empirical knowledge ascertained from decades of studies identified a set of post-trans lational modifications known to provide risk for the developability of a specific 15 molecule. These post-translational modifications have been shown to correlate with amino acid strings present in the primary sequence of the heavy and light chain proteins. Algorithms have been generated that can identify these se quences and determine the potential risk they will have on the manufacturability and developability of a therapeutic antibody.

20 [000164] In silico analysis of the primary sequence of the antibody can be used to de-risk a molecule for its potential to be developed as a therapeutic. In particular, detailed analysis of the VH and VL regions can identified unique amino acids that are deemed important for the molecules activity but also may be a potential risk for its stability over time. Sequence specific 25 deamidation has been identified as a potential risk for protein structures. Protein deamidation can occur on the amide side chains of glutamines or asparagine residues and transform them into a carboxylate group (Lorenzo et al. PLOSone, DOI:10.1371, Dec. (2015)). Nonenzymatic deamidation at neutral pH occurs faster for asparagine and is therefore considered a higher 30 risk than glutamine. The activity is further influenced by the subsequent amino acid in the sequence and can occur at a rate of days or years. The actual fate of the protein that undergoes deamidation needs to be evaluated experimentally to determine the impact of the change both on its stability and activity.

[000165] We identified a site for deamidation within the VH domain of 5 GM37. Amino acid residues 54 is an asparagine(N) followed by a glycine(G) at position 55. The N54 is at high risk for spontaneous deamidation. To mitigate this risk we generated a set of 3 variants that replace the asparagine(N) with serine(S), glutamine(Q) or histidine(H). All 3 variants were produced in mammalian cell culture using transient transfection 10 methods. All 3 variants showed similar expression and purification properties as GM37wt.

[000166] For each of the eight products 400 ml transient transfections were performed using CHOKSV GS-KO cells which had been in culture for minimum 2 weeks. Cells were sub-cultured 24 hours prior to transfection. All 15 transfections were carried out via electroporation using Gene Pulse XCell (Bio-Rad). For each transfection, viable cells were resuspended in pre warmed CD-CHO media supplemented with 6mM L-glutamine to 2.86x107 cells/ml. 40 pg of each established SGV DNA containing the appropriate heavy and light chains were aliquoted into each cuvette (Bio-Rad, 20 GenePulser cuvette, 0.4 cm gap, 165-2088) and 700 pl cell suspension added. Cells were electroporated at 300V, 900pF. Transfected cells were transferred to rep-warmed media in Erlenmeyer flasks and the contents of the cuvettes rinsed twice with prewarmed media were also transferred to the flasks. Transfectant cultures were incubated in a shaking incubator at 36.50C, 25 5%CO2, 85% humidity, 140 rpm for 6 days. Cell viability was measured at the time of harvest using a Cedex HiRes automated cell counter (Rosche).

[000167] In order to evaluate the importance of residue 54 in binding to human alpha-synuclein we analyzed the ability of the variants to bind in two different experiments. Using a competition ELISA format we evaluated the 30 impact the change at residue 54 would have on the ability of GM37 to bind alpha-synuclein in solution. By evaluating the concentration of synuclein able to inhibit binding of the antibody to synuclein coated ELISA plates we showed all three variants maintained the same binding as GM37wt and bind to alpha synuclein with high affinity resulting in IC50s of 1-2nM (Fig. 7). A competition assay was performed using preincubation of a fixed concentration (0.3 pg/ml) 5 of each of the following antibodies, GM37 (named GM 37wt), GM37 variant, GM37 variant2 and GM37 variant3 with a range of 0-1000 nM human alpha synuclein for 60 minutes at room temperature. The remaining unbound antibody was captured and measured on ELISA plates coated with 100 ng/ml of recombinant human alpha-synuclein using an anti-human detection 10 antibody by electrochemiluminesence (MSD, Gathersburg, MD). The IC50s of the interaction are 1.9nM, 1.6nM, 2.1nM and 1.4nM for GM37 wt, GM37variantl, GM37variant2 and GM37variant3, respectively (as determined using Prism Graphpad@).

[000168] Using surface plasmin resonance (SPR), we evaluated the real 15 time kinetics of binding of GM37 wt (2 batches) and the three variants (Example 2). The human alpha-synuclein was captured to the slide(ligand) and the antibodies were each tested at multiple concentrations as analytes. Analysis of the binding curves in the presence of antibody at multiple concentrations showed that the on rates were the same for all four antibodies, 20 similarly when the antibody was removed from the buffer the off-rates measured showed no statistical difference between the antibodies. Using a 1:1 binding algorithm all 4 antibodies have near identical binding constants (Fig. 7). concern over the loss of potency.

25 Example 5: Tau aggregation induced by alpha-synuclein seeds can be pre vented by antibodies against alpha-synuclein

Description of alpha-synuclein fibril (seed) preparation 30 Fibrillation of alpha-synuclein can be done following slightly different protocols.

Recombinant alpha-synuclein purchased from rPeptide (catalog# S-1001-2) was dissolved according to the manufacturers recommendation in double-distilled wa ter resulting in a 1 mg/ml solution in 20 mM Tris-HCL/100 mM NaCl, pH = 7.4. Al pha-synuclein preformed fibrils (PFFs) were generated from monomeric alpha 5 synuclein using the Virginia Lee/Kelvin Luk protocol (Luk et al, Science, 2012, 16;338(6109):949-53). The 1 mg/ml solution was incubated at 37°C with agitation (300 rpm) for 2 days, then a pause for 3 days, then 1 day of agitation, then 1 day pause, then 4 days of agitation. After that the fibrils were harvested and kept at 20°C until use. When fibrils were used in cellular assays, they were always soni 10 cated at 5 min, setting 5.50% cycle, with horn probe sonicator, immediately prior to addition. Alternatively the 1 mg/ml solution is shaken constantly for 5-7 days at 37°C. The end product is termed "crude fibrils". Upon sonication, they are termed "crude seeds". The crude fibrils can be centrifuged and the pellet containing the aggre 15 gated alpha-synuclein is suspended in fresh PBS, and called "pure fibrils". "Pure seeds" are obtained by sonicating pure fibrils.

Alpha-synuclein antibody-mediated inhibition of seeding of Tau 20 In order to show the effect of alpha-synuclein antibody mediated inhibition of seeding of intracellular Tau, a HEK293 cell based seeding assay was setup (Fig ure 8, lower panel illustrates the assay set-up). HEK293 cells were plated 100,000 cells /well in 24 well plates and transiently transfected with cDNA en coding human tau-P301L-FLAG in 24 hours after plating. Twenty four hours after 25 transfection cells where seeded (using Lipofectamine2000 transfection) with ag gregated fibrillated alpha-synculein (seeds) with or without antibodies for 24 hours, followed by splitting and re-plating cells and harvesting after additional 24 hours. Cells were lysed and sonicated in PBS, supplemented with 1% triton X, phos-stop and complete phosphatase and protease inhibitors (Roche) buffer. To 30 tal cell lysates were analyzed using the tau aggregation assay from Cisbio. This assay is based on time-resolved fluorescence using the same antibody for both donor (Tb3+ conjugated) and acceptor (d2 conjugated) antibody in FRET. A 10 pl sample was mixed with 10 pl antibody mix and incubated for 20h. The plate was read on the Pherastar plate reader to assess time-resolved fluorescence (FRET signal measured/integrated after switching of the excitation light). The as say measures aggregated tau both in human brain autopsy material from AD pa 5 tients, brain material from Tau transgenic mice (rTg4510) and in seeded HEK293 cells with high specificity and sensitivity. Using different preparations of fibrillated Tau protein as seed this type of HEK293 cell based seeding assay has been effi ciently used to select Tau antibody clinical candidates. Results are shown in Figure 8, upper panel. Transfection of alpha-synuclein 10 seeds (300ng crude seeds) alone results in a relative tau aggregation around 100 (alpha-Syn, 3 rd bar) indicating that alpha-synuclein potently induce cross seeding of endogenous Tau. The seeding effect was not affected by co-transfec tion with B12 (control antibody) and the following Tau antibodies (mC10-2, mD1.2, an tau binding antibody named LU0041G and humanized (h) C10-2). By 15 co-incubating the cells with four different antibodies against alpha-synuclein HLD1, GM37 (37), GM63 (63) and 9E4 there was however a partial reversal of Tau aggregation, For example the antibody 9E4 results in an increase in relative aggregation to 20 (as compared to 100 in non-treated controls). All antibodies were co-transfected with crude seeds (2.4 ug antibody and 300 ng seeds in a to 20 tal volume of 400ul).

Example 6: Antibody Discovery 2E6 and 2E6 variants

A. Immunization/Hybridoma screening 25 Monoclonal antibodies against alpha-synuclein were generated by immunizing mice with different synuclein aggregates cross linked with for example reactive al dehydes. The first antigen was made of recombinant lyophilized alpha-synuclein from Rpeptide (4241 Mars Hill Road, Bogart, GA 30622, USA). It was made by dissolving the protein in PBS to give a solution of 70 uM alpha-synuclein (1mg/ml). 30 The solution was incubated 18 hours at 37 degrees C and frozen in 100 ul aliquots. The second antigen was made similarly from recombinant alpha-synuclein (Rpep tide) by dissolving it at 70 microM in 20 mM Tris (pH=7.4), 0.15 M NaCl. Reactive aldehyde ONE (4-oxo-2-Nonenal, Cat # 10185 from Cayman Chemicals, Ann Ar bor, MI) was added in a molar ratio of 20:1 to covalently cross link oligomers of alpha-synuclein. The solution was incubated for 18 hours at 37 C (without shaking). The unreacted ONE was removed by Vivaspin500 spin column (10 kDa MWCO) 5 and the samples were dialyzed against 20 mM Tris, pH 7.4, 0.15 M NaCl, and frozen in aliquots. The third antigen was recombinant alpha-synuclein fragment amino acids 1-60 (Rpeptide) which was sent to aslyophilized powder (original ma terial from Rpeptide). Briefly, three female mice (4-7 weeks old) were immunized and boosted up to three times. Tail-bleeds were taken and screened for anti-synu 10 clein antibodies by enzyme-linked immunosorbent assay (ELISA) against the an tigen. Titer is defined by the serum dilutions to achieve OD reading of 3-times the base line in an ELISA. Mice showing a titer greater than 1:50,000 over control were selected for fusion. Harvested splenocytes were fused to SP2/0 mouse myeloma cells, diluted and plated from single cell fusions. Supernatants were harvested 14 15 days post-fusion and screened for antibody production. Using the synuclein ELISA 50 positive clones were recovered from -1000 wells. A Clonotyping System/AP kit was used for immunoglobulin isotyping (Southern Biotechnology, Birmingham, AL). The 50 anti-alpha-synuclein supernatants were screened for reduction of ac cumulation of atto-labelled alpha-synuclein aggregates in the SKMEL5 cell assay 20 below. The commercial antibody LB509 was included as positive control. It was found that out of the 50 antisera, only 4 antisera reduced the intracellular accumu lation of alpha-synuclein and these antibodies were taken forward for cloning. These four antibodies were then tested in dose response in the assay. The anti body with largest effect, 2E6, was selected for further characterization in PD rele 25 vant models.

Description of fibril preparation Recombinant alpha-synuclein was ordered from rPeptide (catalog # S-1001-2) and dissolved according to the manufacturers recommendation in double-distilled 30 water resulting in a 1 mg/ml solution in 20 mM Tris-HCL/100 mM NaCl, pH = 7.4. The alpha-synuclein was fluorescently labelled with Atto488 by using the Atto488 Protein Labeling Kit from Sigma (#38371). A mixture of 30 % Atto488-labelled and 70 % unlabeled alpha-synuclein was made and this mixture was then incu bated at 37°C with agitation (300 rpm) for 2 days, then a pause for 3 days, then 1 day of agitation, then 1 day pause, then 4 days of agitation. After that the fibrils were harvested at kept at -20°C until use. When fibrils were used in cellular as 5 says, they were always sonicated at 5 min, setting 5.50% cycle, with horn probe sonicator, immediately prior to addition.

Antibody-mediated inhibition of accumulation in SK-mel5 cells The human melanoma cell line SK-mel5 (ATCC, HTB-70) was grown in accord 10 ance with the ATCC-guidelines. Cells were plated at a density of 3000 cells per well in Falcon BD 96-well plates and left to adhere overnight. Atto488-labelled al pha-synuclein fibrils were added to the cells (0.01 mg/ml) together with m2E6 antibody (0.01 mg/ml) and alpha-synuclein peptides 113-125 or 126-140 (0.01 mg/ml). After 24 hours of incubation, the cells were washed twice in PBS and 15 fixed by 4 % paraformaldehyde. The cells were then stained with Hoechst and read in Cellomics ArrayScan. Nuclei were detected in one channel and defined the number of valid objects. Atto488-labelled fibrils were detected in another channel in a pre-defined ring-formed area surrounding the nucleus, thus repre senting the cytoplasm of the cells. The percent of the cells containing alpha 20 synuclein spots was quantified. The result shows that in cells not given fibrils, there was only a very low background of spot-containing cells (background were probably due to autofluorescence) Fig 7C. In the cells given fibrils only, 75 % of the cells had accumulated intracellular spots. In the cells co-incubated with fibrils and m2E6 antibody, there were only around 30% spot-positive cells. When the 25 cells were co-incubated with fibrils, m2E6 and the 126-140 peptide, there were around 60% positive cells, thus the peptide significantly inhibited the effect of m2E6. Co-incubation of the 113-120 peptide with fibrils and 2E6 did not change the effect of m2E6. Incubation of fibrils together with either of the peptides 113 120 or 126-140 had no effect on the accumulation of fibrils in the cells. Thus, the 30 m2E6 binds to the alpha-synuclein fibrils in solution and inhibits their accumula tion in the cells.

Treatment with increasing doses of 2E6-HLD1showed a dose-dependent reduc tion in the percentage of cells with spots. Cells treated with irrelevant control anti body (B12) showed no effect.

5 B. Synuclein ELISA Antibody-positive fusions were analysed for binding using an antigen-specific ELISA assay. Corning 96 well high binding plates were coated with 100 ng of ag gregated synuclein. Wells were blocked using 5% milk in PBS for 1 hour (hr) at 10 room temperature (RT). Plates were washed 3 times using PBS+1% Tween 20. One hundred microliters of hybridoma supernatant were added to each well and plates were incubated at RT. Subsequently, HRP-conjugated goat anti-mouse IgG (H&L chain-specific or y-chain specific) secondary was added to each well to de tect the presence of bound anti-synuclein antibody. For quantification substrate, 15 One component TMB, was added and plates were measured at OD620.

C. Determining the DNA sequence of antibody HC and LC variable do mains Four anti-alpha synuclein positive hybridomas were selected and mRNA was ex 20 tracted from cell pellets. cDNAs from each mRNA prep were generated by re verse-transcriptase using oligo(dT) primers. Subsequently, PCR reactions were performed using variable domain primers to amplify both the VH and VL regions of the HC and LC genes. Amplified DNA was separated on an agarose gel and both the VH and VL products were isolated, purified from the gel, cloned into 25 pCR2.1 (Invitrogen) and transformed into TOP10 cells. A minimum of 6 positive colonies were selected and analysed by DNA sequencing to determine the se quence of the VH and VL regions.

Example 7: Antibody Engineering 30 Expression of Monoclonal Antibodies Cultures of hybridoma clones were expanded and mouse monoclonal antibodies were purified from the cultured supernatants using protein G chromatography.

Recombinant mouse, human and chimeric antibodies were produced using tran sient co-transfection of heavy and light chain genes into HEK293 cells, expansion of the cultures, harvesting the supernatants and purification by protein chroma tography. Instances where there was repeated need for gram quantities of anti 5 bodies stable cell lines were created in CHO cells. These stable cell lines could be expanded as needed and antibody purification was performed as before.

Cloning of recombinant antibodies Recombinant monoclonal antibodies were generated by gene synthesis of the 10 heavy and light chain genes (Geneart A/G). Synthesized genes were subse quently cloned into standard expression vectors (e.g. pcDNA3.1) for expression in mammalian cell culture.

Humanization 15 Humanization of m2E6 was carried out by structure based CDR grafting. The amino acid sequences of the 2E6 VL and VH domains were screened for homology against all human antibody VL and VH framework amino acid sequences found in the PDB and IMGT databases. Structural modeling was performed on the m2E6 Fv region using 20SL antibody from the PDB database. The 20SL amino acid 20 sequences are 82.7% and 83.2% homologous to the 2E6 VH and VL domains, respectively. Importantly the structure for 20SL was determined at a resolution of 2.1A. Structural alignment of the 2E6 humanized framework with 20SL enabled determination of important residues in the framework regions that could potentially influence folding or local structure via steric hindrance or steric force. Theoretical 25 antibody structural modeling of the humanized antibody was employed to instruct on the potential importance of maintaining specific residues as the original mouse amino acid in the humanized version of 2E6 in order to maintain binding specificity and affinity. The structural modeling was employed to optimize the activity of hu manized 2E6. 30 Humanization of the 2E6 VH region was performed by grafting the VH CDRs onto the framework of the human germline gene, IGHV1-46*01 (69% homology). There are 23 amino acid differences between the mouse 2E6 and the selected human framework regions. Structural modeling identified 7 amino acid positions where the change to the human residue had the potential to negatively impact the activity of 2E6. These residues were back-mutated to the original mouse amino acids. Three different versions of the humanized heavy chain were produced. Humanized 5 HLD-1 contains all 7 back mutations, M37V, 148M, A68V, L70M, V72R, K74T, A79V, HLD-2 contains 148M, A68V, L70M, V72R, K74T, A79V, and HLD-3 con tains M37V, 148M, L70M, V72R, K74T, A79V.

Humanization of the 2E6 VL region was performed by grafting the VL CDRs onto 10 the framework of the human germline gene, IGKV3-11*01 (64% homology). There are 26 amino acid differences between the mouse 2E6 and the selected human framework regions. Structural modeling identified 4 amino acid positions, R45L, W46L, V571, Y70F, where the change to the human residue had the potential to negatively impact the activity of 2E6. For HLD-1, HLD-2 and HLD-3 all 4 residues 15 were back-mutated to the original mouse amino acids.

HLD-1, HLD-2 and HLD-3 were expressed transiently in HEK 293 cells. Antibodies were purified from cultured supernatants and subsequently analyzed for binding to synuclein by SPR (Biacore 3000) using the synuclein ligand format (Table 5). 20

Table 5: Kinetic binding analysis of different humanized 2E6 clones and chi meric 2E6

ka (1/Ms) kd (1/s) KA (1/M) KD (nM) Chi2 KD improvement

Ch2E6 6,29E+04 2,65E-04 2,38E+08 4,21E-09 3,57 1

HLD1 1,23E+05 2,12E-04 5,81E+08 1,72E-09 4,56 2

HLD2 5,80E+04 2,85E-04 2,04E+08 4,91E-09 4,34 1

HLD3 3 4,89E+04 2,60E-04 1,88E+08 S,32E-09 2,79 1

Affinity maturation of HLD1 was done by randomized mutations in the light chain CDR3 by codon based degenerated PCR primers, and similarly randomized mu tations in the heavy chain CDR3 by codon based degenerated PCR primers and using in vitro evolution with error-prone PCR. Antibodies were purified from cul 5 tured supernatants and subsequently analyzed for binding to synuclein by SPR (Biacore 3000) using IgGs captured using anti human IgG Ab immobilized on the CM5 chip (Table 6).

Table 6: Kinetic binding analysis of different affinity matured versions of hu 10 manized 2E6 clone HLD1 - after first round of affinity maturation

ka (1/Ms) kd (1/s) KA (1/M) KD (M) Chi2 KD improvement

Ch2E6 2.45E+04 1.39E-03 1.76E+07 5.67E-08 0.22 1

HLD1 4.16E+04 9.44E-04 4.40E+07 2.27E-08 0.164 2.5

L3-11 1.45E+05 3.16E-04 4.60E+08 2.18E-09 0.285 26

7A10 5.17E+04 2.85E-04 1.81E+08 5.52E-09 0.297 10.3

9C12 4.95E+04 2.78E-04 1.78E+08 5.62E-09 0.631 10

8D9 7.41E+04 4.83E-04 1.53E+08 6.52E-09 0.301 8.7

7C4 1.23E+05 9.97E-04 1.23E+08 8.12E-09 1.04 7

After first round of affinity maturation we constructed 4 mutations (A, B, C, D): A) combined the two mutations in heavy chain CDR2 (mutate KYNVNFKT to KYNVNIKT) and heavy chain CDR3 (mutate LGHYGNLYAMDY to 15 LGHYGNLYAKDY); B) incorporated light chain CDR1 mutation (mutate SASSSVSYMH to SASSSVSYIH) into the L3-11 light chain; C) incorporated light chain framework mutation (mutate PRRWIY to PRRLlY, immediately upstream CDR2) into the L3-11 light chain; and D) incorporated light chain CDR1 mutation (mutate SASSSVSYMH to SASSSVSYIH) and light chain framework mutation 20 (mutate PRRWIY to PRRLlY) into the L3-11 light chain. Based on the Biacore data and antibody sequence, we tested co-expression of light chain and heavy chain with various combinations: 1. L3-11 light chain + 9C12 heavy chain 2. L3-11 light chain+ 8D9 heavy chain 5 3. 7A10 light chain + 9C12 heavy chain 4. L3-11 light chain+ A 5. 7A10 light chain + A 9. B + 9C12 heavy chain 10. C + 9C12 heavy chain 10 11. D + 9C12 heavy chain 12. B + 8D9 heavy chain 13. C + 8D9 heavy chain 14. D + 8D9 heavy chain 15. B + heavy chain 15 16. C + heavy chain 17. D + heavy chain Antibodies were purified from cultured supernatants and subsequently analyzed for binding to synuclein by SPR (Biacore 3000) using IgGs captured using anti human IgG Ab immobilized on the CM5 chip (Table 7). 20 Table 7: Kinetic binding analysis of different affinity matured versions of hu manized 2E6 clone HLD1 - after combination of mutations KA (1/M) KD (M) Chi2 KD improvement ka (1/Ms) kd (1/s)

Ch2E6 2.45E+04 1.39E-03 1.76E+07 5.67E-08 0.22 1

HLD1 4.16E+04 9.44E-04 4.40E+07 2.27E-08 0.164 2,5

HLDl-14 1.35E+05 5.60E-05 2.42E+09 4.14E-10 0.03 137,0

HLDI-12 2.47E+05 1.12E-04 2.21E+09 4.51E-10 0.12 125,7

HLDI-13 1.46E+05 7.07E-05 2.07E+09 4.83E-10 0.11 117,4

HLD-15 2.5SE+05 1.25E-04 2.06E+09 4.85E-10 0.09 116,9

HLD1-9 2.60E+05 1.33E-04 1.94E+09 5.14E-10 0.06 110,3

HLDl16 L53E+05 8.97E-05 L71E+09 5.85E-10 0A4 96,9

HLDl-2 238E+05 1.52E-04 1.57E+09 6.36E-10 0.06 89,2

HLDI13 9.99E+04 1.26E - 7.94E+08 1.26E-09 OM0 45,0

HLDI-S 9.29E+04 1.28E-04 7.27E+08 1.38E-09 0.03 41,1

Example 8 Frontal cortex from AD patients were homogenized in ice cold sterile PBS and homogenized by a knife homogenizer, sonicated using a Branson sonifier (5 pulses 0,9 seconds, output 2) and cleared at 3000 g, 5 min at 5 4C. Supernatants were collected and protein concentrations determined by BCA. Samples used to determine the level of alpha-synuclein aggre gates contained from 2.1-4.8 ug/pl protein. Alpha-synuclein aggregates was measured using the commercial available alpha-synuclein aggrega tion assay from Cisbio (cat no 6FASYPEG). Alpha-synuclein phosphory 10 lated on serinel29 (synuclein-p129), a marker for Lewy bodies, was meas ured using the upcoming commercial available synuclein-p129 aggrega tion assay from Cisbio. The two assays were performed according to the manufacturers protocols using the respective buffers from each kit. Briefly, all 10% homogenates were serially diluted 1:12 and 1:72 in either of the ly 15 sis buffers and 10 pl sample was mixed with 10 pl of the respective anti body mix (5 pl Tb-cryptate antibody and 5 pl d2 conjugate) and incubated for 20 hours. Time-resolved FRET was measured on the Pherastar plate reader and signal-to-noise ratio was calculated and normalized to protein for each sample. 20 Fifty fresh frozen tissue samples of frontal cortex from AD patients were obtained from the Banner Sun Health Research Institute Brain and Body Donation program (BBDP), Sun city, Arizona, US. 25 samples from pa tients with mid-stage AD (Braak stage Il/IV) and 25 samples from patients 25 with late-stage AD (Braak stage V/VI) was obtained. None of the samples were reported to contain immunohistochemically evidence of alpha-synu clein pathology.

Results are shown in figure 9. Alpha-synuclein aggregates can be meas ured in all 50 AD cases. In figure 9A raw data from 12- and 72- fold dilu tions of 10% brain homogenates (W/V) show a concentration dependent signal intensity. Two samples from patients with Dementia with Lewy bod 5 ies (DLB) were included as positive controls (red lines in figure 9A). In fig ure 9B the data from the 1:12 dilution is normalized to total protein in the samples showing the presence of alpha-synuclein aggregates in similar levels in both mid-stage AD (Braak stage Il/IV) and late-stage AD (Braak stage V/VI). 10 Alpha-synuclein phosphorylated on serine 129 was not detected in any of the AD samples, whereas it was clearly present in the DLB samples (figure 9C). Alpha-synuclein phosphorylated on serine 129 is a marker for mani fest Lewy body pathology and histological staining of post mortem brain 15 with alpha-synuclein-serinel29-phospho antibodies are routinely used to confirm the diagnosis of synucleopathies like Parkinson's disease and DLB. The absence of this marker in the 50 AD samples might indicate that alpha-synuclein aggregates are always present in AD brains - and that synuclein aggregates can be present without the presence of manifest 20 Lewy body pathology.

Based on these findings in combination with the findings in figure 8, we hy pothesize that any alpha-synuclein antibody that are capable of neutraliz ing alpha-synuclein aggregates (seeds) or by other means prevent alpha 25 synuclein aggregates in entering neurons or glia cells and facilitate aggre gation of Tau, will have a therapeutic potential to treat tauopathies.

30

SEQUENCE LISTING

5 <110> H. Lundbeck A/S

<120> Monoclonal Anti-Alpha-Synuclein Antibodies for Prevent-ing

Tau Aggregation

10 <130> 1055

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15 <170> PatentIn version 3.5

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Ala Lys Asn Trp Ala Pro Phe Asp Ser

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Ala Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala

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15 Gly Ala Ser Ser Arg Ala Thr 1 5

<210> 6

20 <211> 9 <212> PRT

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Gln Gln Tyr Gly Ser Ser Pro Trp Thr 30 1 5

<210> 7

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<220> <223> GM37 CDR Heavy Chain

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Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly 1 5 10 15

10 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

15 Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala Ile Arg Ser Asn Gly Asp Arg Thr Asp Tyr Ala Asp Ser Val

20 50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Gln Asn Thr Leu Tyr

65 70 75 80 25

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

30 Ala Lys Asn Trp Ala Pro Phe Asp Ser Trp Gly Gln Gly Thr Leu Val

100 105 110

35 Thr

<210> 8

<211> 108

5 <212> PRT

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<220>

<223> GM 37 Light Chain 10 <400> 8

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

20 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

25 Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 30 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

35

Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

5 <210> 9

<211> 6

<212> PRT <213> Artificial

10 <220>

<223> Epitope 112-117

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15 Ile Leu Glu Asp Met Pro 1 5

<210> 10

20 <211> 140 <212> PRT

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<220> 25 <223> Alpha-synuclein

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Met Asp Val Phe Met Lys Gly Leu Ser Lys Ala Lys Glu Gly Val Val 30 1 5 10 15

Ala Ala Ala Glu Lys Thr Lys Gln Gly Val Ala Glu Ala Ala Gly Lys

20 25 30

35

Thr Lys Glu Gly Val Leu Tyr Val Gly Ser Lys Thr Lys Glu Gly Val

35 40 45

5 Val His Gly Val Ala Thr Val Ala Glu Lys Thr Lys Glu Gln Val Thr 50 55 60

Asn Val Gly Gly Ala Val Val Thr Gly Val Thr Ala Val Ala Gln Lys 10 65 70 75 80

Thr Val Glu Gly Ala Gly Ser Ile Ala Ala Ala Thr Gly Phe Val Lys

85 90 95

15

Lys Asp Gln Leu Gly Lys Asn Glu Glu Gly Ala Pro Gln Glu Gly Ile

100 105 110

20 Leu Glu Asp Met Pro Val Asp Pro Asp Asn Glu Ala Tyr Glu Met Pro

115 120 125

25 Ser Glu Glu Gly Tyr Gln Asp Tyr Glu Pro Glu Ala

130 135 140

<210> 11 30 <211> 165

<212> PRT

<213> Artificial

<220> 35 <223> A-Syn-AAKK-BAP

<400> 11

Met Asp Val Phe Met Lys Gly Leu Ser Lys Ala Lys Glu Gly Val Val

1 5 10 15

5

Ala Ala Ala Glu Lys Thr Lys Gln Gly Val Ala Glu Ala Ala Gly Lys 20 25 30

10 Thr Lys Glu Gly Val Leu Tyr Val Gly Ser Lys Thr Lys Glu Gly Val

35 40 45

15 Val His Gly Val Ala Thr Val Ala Glu Lys Thr Lys Glu Gln Val Thr 50 55 60

Asn Val Gly Gly Ala Val Val Thr Gly Val Thr Ala Val Ala Gln Lys

20 65 70 75 80

Thr Val Glu Gly Ala Gly Asn Ile Ala Ala Ala Thr Gly Leu Val Lys

85 90 95 25

Lys Asp Gln Leu Ala Lys Gln Asn Glu Glu Gly Phe Leu Gln Glu Gly 100 105 110

30 Met Val Asn Asn Thr Asp Ile Pro Val Asp Pro Glu Asn Glu Ala Tyr

115 120 125

35 Glu Met Pro Pro Glu Glu Glu Tyr Gln Asp Tyr Glu Pro Glu Ala Gly 130 135 140

Ser Ala Gly Gly Ser Gly Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys

145 150 155 160

5

Ile Glu Trp His Glu 165

10 <210> 12

<211> 165 <212> PRT

<213> Artificial

15 <220> <223> A-Syn-BAAK-BAP

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20 Met Asp Val Phe Met Lys Gly Leu Ser Met Ala Lys Glu Gly Val Val

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25 Ala Ala Ala Glu Lys Thr Lys Gln Gly Val Thr Glu Ala Ala Glu Lys

20 25 30

Thr Lys Glu Gly Val Leu Tyr Val Gly Ser Lys Thr Lys Glu Gly Val

30 35 40 45

Val His Gly Val Ala Thr Val Ala Glu Lys Thr Lys Glu Gln Val Thr

50 55 60

35

Asn Val Gly Gly Ala Val Val Thr Gly Val Thr Ala Val Ala Gln Lys

65 70 75 80

5 Thr Val Glu Gly Ala Gly Ser Ile Ala Ala Ala Thr Gly Phe Val Lys 85 90 95

Lys Asp Gln Leu Ala Lys Gln Asn Glu Glu Gly Phe Leu Gln Glu Gly 10 100 105 110

Met Val Asn Asn Thr Asp Ile Pro Val Asp Pro Glu Asn Glu Ala Tyr

115 120 125

15

Glu Met Pro Pro Glu Glu Glu Tyr Gln Asp Tyr Glu Pro Glu Ala Gly

130 135 140

20 Ser Ala Gly Gly Ser Gly Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys

145 150 155 160

25 Ile Glu Trp His Glu

165

<210> 13 30 <211> 162

<212> PRT

<213> Artificial

<220> 35 <223> A-Syn-BBAA-BAP

<400> 13

Met Asp Val Phe Met Lys Gly Leu Ser Met Ala Lys Glu Gly Val Val

1 5 10 15

5

Ala Ala Ala Glu Lys Thr Lys Gln Gly Val Thr Glu Ala Ala Glu Lys 20 25 30

10 Thr Lys Glu Gly Val Leu Tyr Val Gly Ser Lys Thr Arg Glu Gly Val

35 40 45

15 Val Gln Gly Val Ala Ser Val Ala Glu Lys Thr Lys Glu Gln Ala Ser 50 55 60

His Leu Gly Gly Ala Val Val Thr Gly Val Thr Ala Val Ala Gln Lys

20 65 70 75 80

Thr Val Glu Gly Ala Gly Ser Ile Ala Ala Ala Thr Gly Phe Val Lys

85 90 95 25

Lys Asp Gln Leu Gly Lys Asn Glu Glu Gly Ala Pro Gln Glu Gly Ile 100 105 110

30 Leu Glu Asp Met Pro Val Asp Pro Asp Asn Glu Ala Tyr Glu Met Pro

115 120 125

35 Ser Glu Glu Gly Tyr Gln Asp Tyr Glu Pro Glu Ala Gly Ser Ala Gly 130 135 140

Gly Ser Gly Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp

145 150 155 160

5

His Glu

10 <210> 14

<211> 165 <212> PRT

<213> Artificial

15 <220> <223> A-Syn-BBKK-BAP

<400> 14

20 Met Asp Val Phe Met Lys Gly Leu Ser Met Ala Lys Glu Gly Val Val

1 5 10 15

25 Ala Ala Ala Glu Lys Thr Lys Gln Gly Val Thr Glu Ala Ala Glu Lys

20 25 30

Thr Lys Glu Gly Val Leu Tyr Val Gly Ser Lys Thr Arg Glu Gly Val

30 35 40 45

Val Gln Gly Val Ala Ser Val Ala Glu Lys Thr Lys Glu Gln Ala Ser

50 55 60

35

His Leu Gly Gly Ala Val Val Thr Gly Val Thr Ala Val Ala Gln Lys

65 70 75 80

5 Thr Val Glu Gly Ala Gly Asn Ile Ala Ala Ala Thr Gly Leu Val Lys 85 90 95

Lys Asp Gln Leu Ala Lys Gln Asn Glu Glu Gly Phe Leu Gln Glu Gly 10 100 105 110

Met Val Asn Asn Thr Asp Ile Pro Val Asp Pro Glu Asn Glu Ala Tyr

115 120 125

15

Glu Met Pro Pro Glu Glu Glu Tyr Gln Asp Tyr Glu Pro Glu Ala Gly

130 135 140

20 Ser Ala Gly Gly Ser Gly Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys

145 150 155 160

25 Ile Glu Trp His Glu

165

<210> 15 30 <211> 141

<212> PRT

<213> Artificial

<220> 35 <223> A-Syn-120-140_Del-BAP

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Met Asp Val Phe Met Lys Gly Leu Ser Lys Ala Lys Glu Gly Val Val

1 5 10 15

5

Ala Ala Ala Glu Lys Thr Lys Gln Gly Val Ala Glu Ala Ala Gly Lys 20 25 30

10 Thr Lys Glu Gly Val Leu Tyr Val Gly Ser Lys Thr Lys Glu Gly Val

35 40 45

15 Val His Gly Val Ala Thr Val Ala Glu Lys Thr Lys Glu Gln Val Thr 50 55 60

Asn Val Gly Gly Ala Val Val Thr Gly Val Thr Ala Val Ala Gln Lys

20 65 70 75 80

Thr Val Glu Gly Ala Gly Ser Ile Ala Ala Ala Thr Gly Phe Val Lys

85 90 95 25

Lys Asp Gln Leu Gly Lys Asn Glu Glu Gly Ala Pro Gln Glu Gly Ile 100 105 110

30 Leu Glu Asp Met Pro Val Asp Gly Ser Ala Gly Gly Ser Gly Gly Leu

115 120 125

35 Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp His Glu 130 135 140

<210> 16

<211> 131

5 <212> PRT

<213> Artificial

<220>

<223> alpha-synuclein amino acids 1-119

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Met Ala His His His His His His Ile Glu Gly Arg Met Asp Val Phe

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15

Met Lys Gly Leu Ser Lys Ala Lys Glu Gly Val Val Ala Ala Ala Glu

20 25 30

20 Lys Thr Lys Gln Gly Val Ala Glu Ala Ala Gly Lys Thr Lys Glu Gly

35 40 45

25 Val Leu Tyr Val Gly Ser Lys Thr Lys Glu Gly Val Val His Gly Val

50 55 60

Ala Thr Val Ala Glu Lys Thr Lys Glu Gln Val Thr Asn Val Gly Gly 30 65 70 75 80

Ala Val Val Thr Gly Val Thr Ala Val Ala Gln Lys Thr Val Glu Gly

85 90 95

35

Ala Gly Ser Ile Ala Ala Ala Thr Gly Phe Val Lys Lys Asp Gln Leu

100 105 110

5 Gly Lys Asn Glu Glu Gly Ala Pro Gln Glu Gly Ile Leu Glu Asp Met 115 120 125

Pro Val Asp 10 130

<210> 17

<211> 106 15 <212> PRT

<213> artificial

<220> <223> kappa (LC constant region)

20 <400> 17

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

1 5 10 15 25

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 20 25 30

30 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

35 40 45

35 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 50 55 60

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

65 70 75 80

5

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 85 90 95

10 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

15 <210> 18

<211> 329 <212> PRT

<213> Artificial

20 <220> <223> IgG1 (HC Constant region)

<400> 18

25 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

30 20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

35

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

5 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 10 85 90 95

Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

15

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

20 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

25 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

30 165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

35

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205

5 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 10 225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

15

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

20 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

25 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 30 305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

35

<210> 19

<211> 4 <212> PRT

<213> Artificial

5 <220>

<223> GM285 epitope 112-115

<400> 19 10 Ile Leu Glu Asp

1

15 <210> 20

<211> 13 <212> PRT

<213> Artificial

20 <220> <223> GM285 CDR1 Heavy Chain

<400> 20

25 Ala Ala Ser Gly Phe Thr Phe Ser Arg Phe Thr Met Thr

1 5 10

<210> 21 30 <211> 17

<212> PRT

<213> Artificial

<220> 35 <223> GM285 CDR2 Heavy Chain

<400> 21

Ala Ile Ser Gly Ser Gly Gly Gly Thr Ser Tyr Ala Asp Ser Val Lys

1 5 10 15

5

Gly

10 <210> 22

<211> 9 <212> PRT

<213> Artificial

15 <220> <223> GM285 CDR3 Heavy Chain

<400> 22

20 Ala Lys Asn Trp Ala Pro Phe Asp Tyr

1 5

25 <210> 23

<211> 12

<212> PRT <213> Artificial

30 <220>

<223> GM285 CDR1 Light Chain

<400> 23

35 Arg Ala Ser Gln Ser Val Ser Arg Ser Tyr Leu Ala 1 5 10

<210> 24

<211> 7

5 <212> PRT

<213> Artificial

<220>

<223> GM285 CDR2 Light Chain 10 <400> 24

Gly Ala Ser Ser Arg Ala Thr

1 5

15

<210> 25

<211> 9 <212> PRT

20 <213> Artificial

<220> <223> GM285 CDR3 Light Chain

25 <400> 25

Gln Gln Tyr Gly Ser Ser Pro Trp Thr 1 5

30 <210> 26

<211> 113 <212> PRT

<213> Artificial

35 <220>

<223> GM285 VH

<400> 26

5 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Phe 10 20 25 30

Thr Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

15

Ser Ala Ile Ser Gly Ser Gly Gly Gly Thr Ser Tyr Ala Asp Ser Val

50 55 60

20 Lys Gly Arg Leu Thr Val Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

25 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Asn Trp Ala Pro Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val

30 100 105 110

Thr

35

<210> 27

<211> 108 <212> PRT

<213> Artificial

5 <220>

<223> GM285 VL

<400> 27 10 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Ser 20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

20 35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60 25

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Val Ser Arg Leu Glu 65 70 75 80

30 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

35 Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105

<210> 28

<211> 329

5 <212> PRT

<213> Artificial

<220>

<223> GM285 IgG1 constant region 10 <400> 28

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

20 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

25 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 30 65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

35

Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

5 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 10 130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

15

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

20 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

25 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

30 210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

35

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

5 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 10 275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

15

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

20 Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

25 <210> 29

<211> 106

<212> PRT <213> Artificial

30 <220>

<223> GM285 Kappa chain

<400> 29

35 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

1 5 10 15

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

20 25 30

5

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 35 40 45

10 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

50 55 60

15 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 65 70 75 80

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

20 85 90 95

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105 25

<210> 30 <211> 113

<212> PRT

30 <213> Artificial

<220> <223> GM37 Variant 1 heavy chain

35 <400> 30

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly

1 5 10 15

5 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 10 35 40 45

Ser Ala Ile Arg Ser Ser Gly Asp Arg Thr Asp Tyr Ala Asp Ser Val

50 55 60

15

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Gln Asn Thr Leu Tyr

65 70 75 80

20 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

25 Ala Lys Asn Trp Ala Pro Phe Asp Ser Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr

30

<210> 31

<211> 113 35 <212> PRT

<213> Artificial

<220> <223> GM 37 variant 2 heavy chain

5 <400> 31

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly 1 5 10 15

10 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

15 Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala Ile Arg Ser Gln Gly Asp Arg Thr Asp Tyr Ala Asp Ser Val

20 50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Gln Asn Thr Leu Tyr

65 70 75 80 25

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

30 Ala Lys Asn Trp Ala Pro Phe Asp Ser Trp Gly Gln Gly Thr Leu Val

100 105 110

35 Thr

<210> 32

<211> 113

5 <212> PRT

<213> Artificial

<220>

<223> GM 37 variant 3 heavy chain 10 <400> 32

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly

1 5 10 15

15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

20 Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

25 Ser Ala Ile Arg Ser His Gly Asp Arg Thr Asp Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Gln Asn Thr Leu Tyr 30 65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

35

Ala Lys Asn Trp Ala Pro Phe Asp Ser Trp Gly Gln Gly Thr Leu Val

100 105 110

5 Thr

<210> 33 10 <211> 17

<212> PRT

<213> Artificial

<220>

15 <223> GM37 variant 1 heavy chain CDR 2

<400> 33

Ala Ile Arg Ser Ser Gly Asp Arg Thr Asp Tyr Ala Asp Ser Val Lys

20 1 5 10 15

Gly

25

<210> 34 <211> 17

<212> PRT

30 <213> Artificial

<220> <223> GM37 variant 2 CDR 2 heavy chain

35 <400> 34

Ala Ile Arg Ser Gln Gly Asp Arg Thr Asp Tyr Ala Asp Ser Val Lys

1 5 10 15

5 Gly

<210> 35 10 <211> 17

<212> PRT

<213> Artificial

<220>

15 <223> GM37 variant 3 CDR 2 heavy chain

<400> 35

Ala Ile Arg Ser His Gly Asp Arg Thr Asp Tyr Ala Asp Ser Val Lys

20 1 5 10 15

Gly

25

<210> 36 <211> 5

<212> PRT

30 <213> Artificial

<220> <223> 9E4 binding epitope

35 <400> 36

Asn Glu Ala Tyr Glu

1 5

5 <210> 37

<211> 134

<212> PRT <213> Artificial

10 <220> <223> HUMAN Beta-synuclein

<400> 37

15 Met Asp Val Phe Met Lys Gly Leu Ser Met Ala Lys Glu Gly Val Val 1 5 10 15

Ala Ala Ala Glu Lys Thr Lys Gln Gly Val Thr Glu Ala Ala Glu Lys

20 20 25 30

Thr Lys Glu Gly Val Leu Tyr Val Gly Ser Lys Thr Arg Glu Gly Val

35 40 45 25

Val Gln Gly Val Ala Ser Val Ala Glu Lys Thr Lys Glu Gln Ala Ser 50 55 60

30 His Leu Gly Gly Ala Val Phe Ser Gly Ala Gly Asn Ile Ala Ala Ala

65 70 75 80

35 Thr Gly Leu Val Lys Arg Glu Glu Phe Pro Thr Asp Leu Lys Pro Glu 85 90 95

Glu Val Ala Gln Glu Ala Ala Glu Glu Pro Leu Ile Glu Pro Leu Met

100 105 110

5

Glu Pro Glu Gly Glu Ser Tyr Glu Asp Pro Pro Gln Glu Glu Tyr Gln 115 120 125

10 Glu Tyr Glu Pro Glu Ala

130

15 <210> 38

<211> 127 <212> PRT

<213> Artificial

20 <220> <223> HUMAN Gamma-synuclein

<400> 38

25 Met Asp Val Phe Lys Lys Gly Phe Ser Ile Ala Lys Glu Gly Val Val

1 5 10 15

Gly Ala Val Glu Lys Thr Lys Gln Gly Val Thr Glu Ala Ala Glu Lys

30 20 25 30

Thr Lys Glu Gly Val Met Tyr Val Gly Ala Lys Thr Lys Glu Asn Val

35 40 45

35

Val Gln Ser Val Thr Ser Val Ala Glu Lys Thr Lys Glu Gln Ala Asn

50 55 60

5 Ala Val Ser Glu Ala Val Val Ser Ser Val Asn Thr Val Ala Thr Lys 65 70 75 80

Thr Val Glu Glu Ala Glu Asn Ile Ala Val Thr Ser Gly Val Val Arg 10 85 90 95

Lys Glu Asp Leu Arg Pro Ser Ala Pro Gln Gln Glu Gly Glu Ala Ser

100 105 110

15

Lys Glu Lys Glu Glu Val Ala Glu Glu Ala Gln Ser Gly Gly Asp

115 120 125

20 <210> 39

<211> 140 <212> PRT

<213> Artificial 25 <220>

<223> alpha-synuclein ortholog for Cynomolgus monkey

<400> 39

30 Met Asp Val Phe Met Lys Gly Leu Ser Lys Ala Lys Glu Gly Val Val

1 5 10 15

35 Ala Ala Ala Glu Lys Thr Lys Gln Gly Val Ala Glu Ala Ala Gly Lys 20 25 30

Thr Lys Glu Gly Val Leu Tyr Val Gly Ser Lys Thr Lys Glu Gly Val

35 40 45

5

Val His Gly Val Ala Thr Val Ala Glu Lys Thr Lys Glu Gln Val Thr 50 55 60

10 Asn Val Gly Gly Ala Val Val Thr Gly Val Thr Ala Val Ala Gln Lys

65 70 75 80

15 Thr Val Glu Gly Ala Gly Ser Ile Ala Ala Ala Thr Gly Phe Ile Lys 85 90 95

Lys Asp Gln Leu Gly Lys Asn Glu Glu Gly Ala Pro Gln Glu Gly Ile

20 100 105 110

Leu Gln Asp Met Pro Val Asp Pro Asp Asn Glu Ala Tyr Glu Met Pro

115 120 125 25

Ser Glu Glu Gly Tyr Gln Asp Tyr Glu Pro Glu Ala 130 135 140

30 <210> 40

<211> 140 <212> PRT

<213> Artificial

35 <220>

<223> alpha-synuclein ortholog for Rat

<400> 40

5 Met Asp Val Phe Met Lys Gly Leu Ser Lys Ala Lys Glu Gly Val Val 1 5 10 15

Ala Ala Ala Glu Lys Thr Lys Gln Gly Val Ala Glu Ala Ala Gly Lys 10 20 25 30

Thr Lys Glu Gly Val Leu Tyr Val Gly Ser Lys Thr Lys Glu Gly Val

35 40 45

15

Val His Gly Val Thr Thr Val Ala Glu Lys Thr Lys Glu Gln Val Thr

50 55 60

20 Asn Val Gly Gly Ala Val Val Thr Gly Val Thr Ala Val Ala Gln Lys

65 70 75 80

25 Thr Val Glu Gly Ala Gly Asn Ile Ala Ala Ala Thr Gly Phe Val Lys

85 90 95

Lys Asp Gln Met Gly Lys Gly Glu Glu Gly Tyr Pro Gln Glu Gly Ile

30 100 105 110

Leu Glu Asp Met Pro Val Asp Pro Ser Ser Glu Ala Tyr Glu Met Pro

115 120 125

35

Ser Glu Glu Gly Tyr Gln Asp Tyr Glu Pro Glu Ala 130 135 140

5 <210> 41

<211> 140

<212> PRT <213> Artificial

10 <220>

<223> alpha-synuclein ortholog for Mouse

<400> 41

15 Met Asp Val Phe Met Lys Gly Leu Ser Lys Ala Lys Glu Gly Val Val 1 5 10 15

Ala Ala Ala Glu Lys Thr Lys Gln Gly Val Ala Glu Ala Ala Gly Lys

20 20 25 30

Thr Lys Glu Gly Val Leu Tyr Val Gly Ser Lys Thr Lys Glu Gly Val

35 40 45 25

Val His Gly Val Thr Thr Val Ala Glu Lys Thr Lys Glu Gln Val Thr 50 55 60

30 Asn Val Gly Gly Ala Val Val Thr Gly Val Thr Ala Val Ala Gln Lys

65 70 75 80

35 Thr Val Glu Gly Ala Gly Asn Ile Ala Ala Ala Thr Gly Phe Val Lys 85 90 95

Lys Asp Gln Met Gly Lys Gly Glu Glu Gly Tyr Pro Gln Glu Gly Ile

100 105 110

5

Leu Glu Asp Met Pro Val Asp Pro Gly Ser Glu Ala Tyr Glu Met Pro 115 120 125

10 Ser Glu Glu Gly Tyr Gln Asp Tyr Glu Pro Glu Ala

130 135 140

15 <210> 42

<211> 446 <212> PRT

<213> artificial

20 <220> <223> 9E4 HC

<400> 42

25 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

30 20 25 30

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

35

Ala Ser Ile Ser Ser Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Asn Val

50 55 60

5 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 10 85 90 95

Ala Arg Gly Gly Ala Gly Ile Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

15

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

20 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

25 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

30 165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

35

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

5 Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe 10 225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

15

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

20 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

25 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 30 305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

35

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

5 Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 10 370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

15

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

20 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

25 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 43 30 <211> 220

<212> PRT

<213> artificial

<220> 35 <223> 9E4 LC

<400> 43

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

5

Asp Arg Val Thr Ile Thr Cys Lys Ser Ile Gln Thr Leu Leu Tyr Ser 20 25 30

10 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Phe Gln Gln Lys Pro Gly Lys

35 40 45

15 Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Ile Arg Lys Ser Gly Val 50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

20 65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Leu Ala Thr Tyr Tyr Cys Gln Gln

85 90 95 25

Tyr Tyr Ser Tyr Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile 100 105 110

30 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

35 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 145 150 155 160

5

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 165 170 175

10 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

15 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

20 210 215 220

<210> 44

<211> 10 25 <212> PRT

<213> artificial

<220>

<223> 9E4 CDR1 Heavy Chain

30 <400> 44

Gly Phe Thr Phe Ser Asn Tyr Gly Met Ser

1 5 10

35

<210> 45

<211> 17 <212> PRT

<213> artificial

5 <220>

<223> 9E4 CDR2 Heavy Chain

<400> 45 10 Ser Ile Ser Ser Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Asn Val Lys

1 5 10 15

15 Gly

<210> 46

20 <211> 7 <212> PRT

<213> artificial

<220> 25 <223> 9E4 CDR3 Heavy Chain

<400> 46

Gly Gly Ala Gly Ile Asp Tyr 30 1 5

<210> 47

<211> 17 35 <212> PRT

<213> artificial

<220> <223> 9E4 CDR1 Light Chain

5 <400> 47

Lys Ser Ile Gln Thr Leu Leu Tyr Ser Ser Asn Gln Lys Asn Tyr Leu 1 5 10 15

10 Ala

15 <210> 48

<211> 7 <212> PRT

<213> artificial

20 <220> <223> 9E4 CDR2 Light Chain

<400> 48

25 Trp Ala Ser Ile Arg Lys Ser

1 5

<210> 49 30 <211> 9

<212> PRT

<213> artificial

<220> 35 <223> 9E4 CDR3 Light Chain

<400> 49

Gln Gln Tyr Tyr Ser Tyr Pro Leu Thr

1 5

5

<210> 50 <211> 13

<212> PRT 10 <213> artificial

<220> <223> GM63 Epitope 126-138

15 <400> 50

Glu Met Pro Ser Glu Glu Gly Tyr Gln Asp Tyr Glu Pro

1 5 10

20 <210> 51

<211> 13 <212> PRT

<213> artificial 25 <220>

<223> GM63 CDR1 Heavy Chain

<400> 51

30 Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr Gly Ile Ile

1 5 10

35 <210> 52

<211> 10

<212> PRT

<213> artificial

<220> 5 <223> GM63 CDR2 Heavy Chain

<400> 52

Trp Ile Ser Ala Tyr Asn Gly Lys Thr Asn 10 1 5 10

<210> 53

<211> 10 15 <212> PRT

<213> artificial

<220> <223> GM63 CDR3 Heavy Chain

20 <400> 53

Thr Arg Ala His Trp Gly Arg Phe Asp Tyr

1 5 10 25

<210> 54 <211> 11

<212> PRT

30 <213> artificial

<220> <223> GM63 CDR1 Light Chain

35 <400> 54

Arg Ala Ser Gln Gly Ile Ser Ser Ala Leu Ala

1 5 10

5 <210> 55

<211> 8

<212> PRT <213> artificial

10 <220>

<223> GM63 CDR2 Light Chain

<400> 55

15 Tyr Asp Ala Ser Ser Leu Glu Ser 1 5

<210> 56

20 <211> 9 <212> PRT

<213> artificial

<220> 25 <223> GM63 CDR3 Light Chain

<400> 56

Gln Gln Phe Lys Ser Tyr Pro Arg Thr 30 1 5

<210> 57

<211> 114 35 <212> PRT

<213> artificial

<220> <223> GM63 VH

5 <400> 57

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15

10 Ser Val Ile Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

15 Gly Ile Ile Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45

Gly Trp Ile Ser Ala Tyr Asn Gly Lys Thr Asn Tyr Ala Gln Asn Leu

20 50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80 25

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Met Tyr Tyr Cys 85 90 95

30 Thr Arg Ala His Trp Gly Arg Phe Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

35 Val Thr

<210> 58

<211> 107

5 <212> PRT

<213> artificial

<220>

<223> GM63 VL 10 <400> 58

Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30

20 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

25 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 30 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Lys Ser Tyr Pro Arg

85 90 95

35

Thr Leu Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

5 <210> 59

<211> 701

<212> PRT <213> artificial

10 <220>

<223> GM63 Heavy CHain Constant Domain

<400> 59

15 Gly Cys Cys Thr Cys Cys Ala Cys Cys Ala Ala Gly Gly Gly Cys Cys 1 5 10 15

Cys Ala Thr Cys Gly Gly Thr Cys Thr Thr Cys Cys Cys Ala Cys Thr

20 20 25 30

Gly Gly Cys Gly Cys Cys Cys Thr Cys Cys Thr Cys Cys Ala Ala Gly

35 40 45 25

Ala Gly Cys Ala Cys Cys Ala Gly Cys Gly Gly Cys Gly Gly Cys Ala 50 55 60

30 Cys Ala Gly Cys Cys Gly Cys Cys Cys Thr Gly Gly Gly Cys Thr Gly

65 70 75 80

35 Cys Cys Thr Gly Gly Thr Gly Ala Ala Gly Gly Ala Cys Thr Ala Cys 85 90 95

Thr Thr Cys Cys Cys Cys Gly Ala Gly Cys Cys Ala Gly Thr Gly Ala

100 105 110

5

Cys Cys Gly Thr Gly Thr Cys Cys Thr Gly Gly Ala Ala Cys Thr Cys 115 120 125

10 Thr Gly Gly Cys Gly Cys Cys Cys Thr Gly Ala Cys Cys Thr Cys Cys

130 135 140

15 Gly Gly Cys Gly Thr Gly Cys Ala Cys Ala Cys Cys Thr Thr Cys Cys 145 150 155 160

Cys Cys Gly Cys Cys Gly Thr Gly Cys Thr Gly Cys Ala Gly Ala Gly

20 165 170 175

Cys Ala Gly Cys Gly Gly Cys Cys Thr Gly Thr Ala Cys Ala Gly Cys

180 185 190 25

Cys Thr Gly Ala Gly Cys Ala Gly Cys Gly Thr Gly Gly Thr Gly Ala 195 200 205

30 Cys Cys Gly Thr Gly Cys Cys Cys Ala Gly Cys Ala Gly Cys Ala Gly

210 215 220

35 Cys Cys Thr Gly Gly Gly Cys Ala Cys Cys Cys Ala Gly Ala Cys Cys 225 230 235 240

Thr Ala Cys Ala Thr Cys Thr Gly Cys Ala Ala Cys Gly Thr Gly Ala

245 250 255

5

Ala Cys Cys Ala Cys Ala Ala Gly Cys Cys Cys Ala Gly Cys Ala Ala 260 265 270

10 Cys Ala Cys Cys Ala Ala Gly Gly Thr Gly Gly Ala Cys Ala Ala Gly

275 280 285

15 Ala Gly Ala Gly Thr Gly Gly Ala Gly Cys Cys Cys Ala Ala Gly Ala 290 295 300

Gly Cys Thr Gly Cys Gly Ala Cys Ala Ala Gly Ala Cys Cys Cys Ala

20 305 310 315 320

Cys Ala Cys Cys Thr Gly Cys Cys Cys Cys Cys Cys Cys Thr Gly Cys

325 330 335 25

Cys Cys Ala Gly Cys Cys Cys Cys Ala Gly Ala Gly Cys Thr Gly Cys 340 345 350

30 Thr Gly Gly Gly Cys Gly Gly Ala Cys Cys Cys Ala Gly Cys Gly Thr

355 360 365

35 Gly Thr Thr Cys Cys Thr Gly Thr Thr Cys Cys Cys Cys Cys Cys Cys 370 375 380

Ala Ala Gly Cys Cys Cys Ala Ala Gly Gly Ala Cys Ala Cys Cys Cys

385 390 395 400

5

Thr Gly Ala Thr Gly Ala Thr Cys Ala Gly Cys Ala Gly Gly Ala Cys 405 410 415

10 Cys Cys Cys Cys Gly Ala Gly Gly Thr Gly Ala Cys Cys Thr Gly Cys

420 425 430

15 Gly Thr Gly Gly Thr Gly Gly Thr Gly Gly Ala Cys Gly Thr Gly Ala 435 440 445

Gly Cys Cys Ala Cys Gly Ala Gly Gly Ala Cys Cys Cys Ala Gly Ala

20 450 455 460

Gly Gly Thr Gly Ala Ala Gly Thr Thr Cys Ala Ala Cys Thr Gly Gly

465 470 475 480 25

Thr Ala Cys Gly Thr Gly Gly Ala Cys Gly Gly Cys Gly Thr Gly Gly 485 490 495

30 Ala Gly Gly Thr Gly Cys Ala Cys Ala Ala Cys Gly Cys Cys Ala Ala

500 505 510

35 Gly Ala Cys Cys Ala Ala Gly Cys Cys Cys Ala Gly Ala Gly Ala Gly 515 520 525

Gly Ala Gly Cys Ala Gly Thr Ala Cys Ala Ala Cys Ala Gly Cys Ala

530 535 540

5

Cys Cys Thr Ala Cys Ala Gly Gly Gly Thr Gly Gly Thr Gly Thr Cys 545 550 555 560

10 Cys Gly Thr Gly Cys Thr Gly Ala Cys Cys Gly Thr Gly Cys Thr Gly

565 570 575

15 Cys Ala Cys Cys Ala Gly Gly Ala Cys Thr Gly Gly Cys Thr Gly Ala 580 585 590

Ala Cys Gly Gly Cys Ala Ala Gly Gly Ala Ala Thr Ala Cys Ala Ala

20 595 600 605

Gly Thr Gly Cys Ala Ala Gly Gly Thr Cys Thr Cys Cys Ala Ala Cys

610 615 620 25

Ala Ala Gly Gly Cys Cys Cys Thr Gly Cys Cys Ala Gly Cys Cys Cys 625 630 635 640

30 Cys Cys Ala Thr Cys Gly Ala Ala Ala Ala Gly Ala Cys Cys Ala Thr

645 650 655

35 Cys Ala Gly Cys Ala Ala Gly Gly Cys Cys Ala Ala Gly Gly Gly Cys 660 665 670

Cys Ala Gly Cys Cys Ala Cys Gly Gly Gly Ala Gly Cys Cys Cys Cys

675 680 685

5

Ala Gly Gly Thr Gly Thr Ala Cys Ala Cys Cys Cys Thr 690 695 700

10 <210> 60

<211> 323 <212> PRT

<213> artificial

15 <220> <223> GM63 Kappa Light Chain Constant Domain

<400> 60

20 Gly Ala Ala Cys Thr Gly Thr Gly Gly Cys Thr Gly Cys Ala Cys Cys

1 5 10 15

25 Ala Thr Cys Thr Gly Thr Cys Thr Thr Cys Ala Thr Cys Thr Thr Cys

20 25 30

Cys Cys Gly Cys Cys Ala Thr Cys Thr Gly Ala Thr Gly Ala Gly Cys

30 35 40 45

Ala Gly Thr Thr Gly Ala Ala Ala Thr Cys Thr Gly Gly Ala Ala Cys

50 55 60

35

Thr Gly Cys Cys Thr Cys Ala Gly Thr Gly Gly Thr Gly Thr Gly Cys

65 70 75 80

5 Cys Thr Gly Cys Thr Gly Ala Ala Cys Ala Ala Cys Thr Thr Cys Thr 85 90 95

Ala Cys Cys Cys Cys Cys Gly Gly Gly Ala Gly Gly Cys Cys Ala Ala 10 100 105 110

Gly Gly Thr Gly Cys Ala Gly Thr Gly Gly Ala Ala Gly Gly Thr Gly

115 120 125

15

Gly Ala Cys Ala Ala Cys Gly Cys Cys Cys Thr Gly Cys Ala Gly Ala

130 135 140

20 Gly Cys Gly Gly Cys Ala Ala Cys Ala Gly Cys Cys Ala Gly Gly Ala

145 150 155 160

25 Gly Ala Gly Cys Gly Thr Cys Ala Cys Cys Gly Ala Gly Cys Ala Gly

165 170 175

Gly Ala Cys Ala Gly Cys Ala Ala Gly Gly Ala Cys Thr Cys Cys Ala

30 180 185 190

Cys Cys Thr Ala Cys Ala Gly Cys Cys Thr Gly Ala Gly Cys Ala Gly

195 200 205

35

Cys Ala Cys Cys Cys Thr Gly Ala Cys Cys Cys Thr Gly Thr Cys Cys

210 215 220

5 Ala Ala Gly Gly Cys Cys Gly Ala Cys Thr Ala Cys Gly Ala Gly Ala 225 230 235 240

Ala Gly Cys Ala Cys Ala Ala Gly Gly Thr Gly Thr Ala Cys Gly Cys 10 245 250 255

Cys Thr Gly Cys Gly Ala Gly Gly Thr Gly Ala Cys Cys Cys Ala Cys

260 265 270

15

Cys Ala Gly Gly Gly Cys Cys Thr Gly Thr Cys Cys Ala Gly Cys Cys

275 280 285

20 Cys Cys Gly Thr Gly Ala Cys Cys Ala Ala Gly Ala Gly Cys Thr Thr

290 295 300

25 Cys Ala Ala Cys Ala Gly Gly Gly Gly Cys Gly Ala Gly Thr Gly Cys

305 310 315 320

Thr Gly Ala

30

<210> 61

<211> 15 35 <212> PRT

<213> artificial

<220> <223> 9E4 Epitope 126-140

5 <400> 61

Glu Met Pro Ser Glu Glu Gly Tyr Gln Asp Tyr Glu Pro Glu Ala 1 5 10 15

10 <210> 62

<211> 5 <212> PRT

<213> artificial

15 <220> <223> CDR1 VH

<400> 62

20 Ser Tyr Trp Met His

1 5

25 <210> 63

<211> 17

<212> PRT <213> artificial

30 <220>

<223> CDR2 VH

<400> 63

35 Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe Lys 1 5 10 15

Thr

5

<210> 64 <211> 12

<212> PRT 10 <213> artificial

<220> <223> CDR3 VH

15 <400> 64

Leu Gly His Tyr Gly Asn Leu Tyr Ala Met Asp Tyr

1 5 10

20 <210> 65

<211> 10 <212> PRT

<213> artificial 25 <220>

<223> CDR1 VL

<400> 65

30 Ser Ala Ser Ser Ser Val Ser Tyr Met His

1 5 10

35 <210> 66

<211> 7

<212> PRT

<213> artificial

<220>

5 <223> CDR2 VL

<400> 66

Asp Thr Ser Lys Leu Ala Ser

10 1 5

<210> 67

<211> 9 15 <212> PRT

<213> artificial

<220> <223> CDR3 VL

20 <400> 67

Gln Gln Trp Ser Ser Asn Pro Pro Thr

1 5 25

<210> 68 <211> 118

<212> PRT

30 <213> artificial

<220> <223> m2E6 VH

35 <400> 68

Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala

1 5 10 15

5 Ser Vai Lys Leu Ser Cys Thr Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Met His Trp Met Lys Gln Arg Pro Gly Arg Gly Leu Glu Trp Ile 10 35 40 45

Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe

50 55 60

15

Lys Thr Lys Ala Thr Leu Thr Val Asp Lys Pro Ser Ser Thr Ala Tyr

65 70 75 80

20 Met His Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

25 Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Met Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Ser Val Thr

30 115

<210> 69

<211> 106 35 <212> PRT

<213> artificial

<220> <223> m2E6 VL

5 <400> 69

Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly 1 5 10 15

10 Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

20 25 30

15 His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr 35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Thr Arg Phe Ser Gly Ser

20 50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Asp Thr Glu

65 70 75 80 25

Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr 85 90 95

30 Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

100 105

35 <210> 70

<211> 118

<212> PRT

<213> artificial

<220> 5 <223> ch2E6 VH

<400> 70

Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala 10 1 5 10 15

Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

15

Trp Met His Trp Met Lys Gln Arg Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

20 Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe

50 55 60

25 Lys Thr Lys Ala Thr Leu Thr Val Asp Lys Pro Ser Ser Thr Ala Tyr

65 70 75 80

Met His Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

30 85 90 95

Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Met Asp Tyr Trp Gly

100 105 110

35

Gln Gly Thr Ser Val Thr

115

5 <210> 71

<211> 106

<212> PRT <213> artificial

10 <220> <223> ch2E6 VL

<400> 71

15 Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly 1 5 10 15

Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

20 20 25 30

His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr

35 40 45 25

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Thr Arg Phe Ser Gly Ser 50 55 60

30 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Asp Thr Glu

65 70 75 80

35 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr 85 90 95

Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

100 105

5

<210> 72 <211> 118

<212> PRT 10 <213> artificial

<220> <223> 2E6-HLD1 VH

15 <400> 72

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

20 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

25 Trp Met His Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe

30 50 55 60

Lys Thr Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

35

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

5 Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Met Asp Tyr Trp Gly 100 105 110

Gln Gly Thr Leu Val Thr 10 115

<210> 73

<211> 107 15 <212> PRT

<213> artificial

<220> <223> 2E6-HLD1 VL

20 <400> 73

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15 25

Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30

30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

35 Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu

65 70 75 80

5

Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr 85 90 95

10 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg

100 105

15 <210> 74

<211> 118 <212> PRT

<213> Artificial

20 <220> <223> 2E6-HLD2 VH

<400> 74

25 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

30 20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

35

Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe

50 55 60

5 Lys Thr Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 10 85 90 95

Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Met Asp Tyr Trp Gly

100 105 110

15

Gln Gly Thr Leu Val Thr

115

20 <210> 75

<211> 107 <212> PRT

<213> artificial 25 <220>

<223> 2E6-HLD2 VL

<400> 75

30 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

35 Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr 35 40 45

5

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60

10 Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu

65 70 75 80

15 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr 85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg

20 100 105

<210> 76

<211> 118 25 <212> PRT

<213> artificial

<220>

<223> 2E6-HLD 3 VH

30 <400> 76

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

35

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

5 Trp Met His Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45

Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe 10 50 55 60

Lys Thr Arg Val Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

15

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

20 Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Met Asp Tyr Trp Gly

100 105 110

25 Gln Gly Thr Leu Val Thr

115

<210> 77 30 <211> 107

<212> PRT

<213> artificial

<220> 35 <223> 2E6-HLD 3 VL

<400> 77

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

5

Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30

10 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

15 Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu

20 65 70 75 80

Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Asn

85 90 95 25

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105

30 <210> 78

<211> 16 <212> PRT

<213> Artificial

35 <220>

<223> D1.2 CDR 1 Light Chain

<400> 78

5 Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr Leu His 1 5 10 15

<210> 79

10 <211> 7

<212> PRT

<213> Artificial

<220> 15 <223> D1.2 CDR 2 Light Chain

<400> 79

Lys Val Ser Asn Arg Phe Ser

20 1 5

<210> 80

<211> 7 25 <212> PRT

<213> Artificial

<220>

<223> D1.2 CDR 3 Light Chain

30 <400> 80

Ser Gln Ser Thr His Val Pro

1 5

35

<210> 81

<211> 13 <212> PRT

<213> Artificial

5 <220>

<223> D1.2 CDR 1 Heavy Chaiin

<400> 81 10 Lys Ala Ser Gly Asn Thr Phe Thr Asp Tyr Glu Ile His

1 5 10

15 <210> 82

<211> 17 <212> PRT

<213> Artificial

20 <220> <223> D1.2 CDR 2 Heavy Chain

<400> 82

25 Ala Ile Asp Pro Glu Thr Gly Asn Thr Ala Tyr Asn Gln Lys Phe Lys

1 5 10 15

Gly

30

<210> 83

<211> 6

35 <212> PRT

<213> Artificial

<220> <223> D1.2 CDR 3 Heavy Chain

5 <400> 83

Ser Arg Gly Phe Asp Tyr 1 5

10 <210> 84

<211> 219 <212> PRT

<213> Artificial

15 <220> <223> D1.2 Light Chain

<400> 84

20 Asp Val Met Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly

1 5 10 15

25 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser

20 25 30

Asn Gly Asn Thr Tyr Leu His Trp His Leu Gln Lys Pro Gly Gln Ser

30 35 40 45

Pro Lys Phe Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

35

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

5 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser 85 90 95

Thr His Val Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 10 100 105 110

Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu

115 120 125

15

Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe

130 135 140

20 Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg

145 150 155 160

25 Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser

165 170 175

Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu

30 180 185 190

Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser

195 200 205

35

Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys

210 215

5 <210> 85

<211> 451

<212> PRT <213> Artificial

10 <220>

<223> D1.2 Heavy Chain

<400> 85

15 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ala 1 5 10 15

Ser Val Thr Leu Ser Cys Lys Ala Ser Gly Asn Thr Phe Thr Asp Tyr

20 20 25 30

Glu Ile His Trp Val Lys Gln Thr Pro Val His Gly Leu Glu Trp Ile

35 40 45 25

Gly Ala Ile Asp Pro Glu Thr Gly Asn Thr Ala Tyr Asn Gln Lys Phe 50 55 60

30 Lys Gly Lys Ala Arg Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

35 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Thr Arg Ser Arg Gly Phe Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr

100 105 110

5

Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro Leu Ala Pro

115 120 125

10 Gly Cys Gly Asp Thr Thr Gly Ser Ser Val Thr Leu Gly Cys Leu Val

130 135 140

15 Lys Gly Tyr Phe Pro Glu Ser Val Thr Val Thr Trp Asn Ser Gly Ser 145 150 155 160

Leu Ser Ser Ser Val His Thr Phe Pro Ala Leu Leu Gln Ser Gly Leu

20 165 170 175

Tyr Thr Met Ser Ser Ser Val Thr Val Pro Ser Ser Thr Trp Pro Ser

180 185 190 25

Gln Thr Val Thr Cys Ser Val Ala His Pro Ala Ser Ser Thr Thr Val 195 200 205

30 Asp Lys Lys Leu Glu Pro Ser Gly Pro Ile Ser Thr Ile Asn Pro Cys

210 215 220

35 Pro Pro Cys Lys Glu Cys His Lys Cys Pro Ala Pro Asn Leu Glu Gly 225 230 235 240

Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Ile Lys Asp Val Leu Met

245 250 255

5

Ile Ser Leu Thr Pro Lys Val Thr Cys Val Val Val Asp Val Ser Glu 260 265 270

10 Asp Asp Pro Asp Val Arg Ile Ser Trp Phe Val Asn Asn Val Glu Val

275 280 285

15 His Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr Asn Ser Thr Ile 290 295 300

Arg Val Val Ser Ala Leu Pro Ile Gln His Gln Asp Trp Met Ser Gly

20 305 310 315 320

Lys Glu Phe Lys Cys Lys Val Asn Asn Lys Asp Leu Pro Ser Pro Ile

325 330 335 25

Glu Arg Thr Ile Ser Lys Ile Lys Gly Leu Val Arg Ala Pro Gln Val 340 345 350

30 Tyr Ile Leu Pro Pro Pro Ala Glu Gln Leu Ser Arg Lys Asp Val Ser

355 360 365

35 Leu Thr Cys Leu Val Val Gly Phe Asn Pro Gly Asp Ile Ser Val Glu 370 375 380

Trp Thr Ser Asn Gly His Thr Glu Glu Asn Tyr Lys Asp Thr Ala Pro

385 390 395 400

5

Val Leu Asp Ser Asp Gly Ser Tyr Phe Ile Tyr Ser Lys Leu Asp Ile 405 410 415

10 Lys Thr Ser Lys Trp Glu Lys Thr Asp Ser Phe Ser Cys Asn Val Arg

420 425 430

15 His Glu Gly Leu Lys Asn Tyr Tyr Leu Lys Lys Thr Ile Ser Arg Ser 435 440 445

Pro Gly Lys

20 450

<210> 86

<211> 11 25 <212> PRT

<213> Artificial

<220>

<223> C10.2 CDR 1 Light Chain

30 <400> 86

Gln Ala Ser Gln Gly Thr Ser Ile Asn Leu Asn

1 5 10

35

<210> 87

<211> 7 <212> PRT

<213> Artificial

5 <220>

<223> C10.2 CDR 2 Light Chain

<400> 87 10 Gly Ala Ser Asn Leu Glu Asp

1 5

15 <210> 88

<211> 7 <212> PRT

<213> Artificial

20 <220> <223> C10.2 CDR 3 Light Chain

<400> 88

25 Leu Gln His Thr Tyr Leu Pro

1 5

<210> 89 30 <211> 13

<212> PRT

<213> Artificial

<220> 35 <223> C10.2 CDR 1 Heavy Chain

<400> 89

Lys Ala Ser Gly Tyr Thr Phe Thr Asp Arg Thr Ile His

1 5 10

5

<210> 90 <211> 17

<212> PRT 10 <213> Artificial

<220> <223> C10.2 CDR 2 Heavy Chain

15 <400> 90

Tyr Ile Tyr Pro Gly Asp Gly Ser Thr Lys Tyr Asn Glu Asn Phe Lys

1 5 10 15

20 Gly

25 <210> 91

<211> 6

<212> PRT <213> Artificial

30 <220>

<223> C10.2 CDR 3 Heary Chain

<400> 91

35 Arg Gly Ala Met Asp Tyr 1 5

<210> 92

<211> 214

5 <212> PRT

<213> Artificial

<220>

<223> C10.2 Light Chain 10 <400> 92

Asp Val Gln Met Ile Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly

1 5 10 15

15

Asp Ile Val Thr Met Thr Cys Gln Ala Ser Gln Gly Thr Ser Ile Asn

20 25 30

20 Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

25 Tyr Gly Ala Ser Asn Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Arg Tyr Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Asp 30 65 70 75 80

Glu Asp Met Ala Thr Tyr Phe Cys Leu Gln His Thr Tyr Leu Pro Phe

85 90 95

35

Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala Ala

100 105 110

5 Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu Thr Ser Gly 115 120 125

Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile 10 130 135 140

Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn Gly Val Leu

145 150 155 160

15

Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser

165 170 175

20 Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Ser Tyr

180 185 190

25 Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser

195 200 205

Phe Asn Arg Asn Glu Cys

30 210

<210> 93

<211> 439 35 <212> PRT

<213> Artificial

<220> <223> C10.2 Heavy Chain

5 <400> 93

Gln Val Gln Leu Gln Gln Ser Asp Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15

10 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Arg

20 25 30

15 Thr Ile His Trp Val Lys Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile 35 40 45

Gly Tyr Ile Tyr Pro Gly Asp Gly Ser Thr Lys Tyr Asn Glu Asn Phe

20 50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80 25

Met Gln Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95

30 Ala Arg Arg Gly Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr

100 105 110

35 Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro Leu Ala Pro 115 120 125

Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly Cys Leu Val

130 135 140

5

Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn Ser Gly Ser 145 150 155 160

10 Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Asp Leu

165 170 175

15 Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser Ser Thr Trp Pro Ser

180 185 190

Glu Thr Val Thr Cys Asn Val Ala His Pro Ala Ser Ser Thr Lys Val

20 195 200 205

Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys Lys Pro Cys Ile Cys

210 215 220 25

Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe Pro Pro Lys Pro Lys 225 230 235 240

30 Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val Thr Cys Val Val Val

245 250 255

35 Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe Ser Trp Phe Val Asp 260 265 270

Asp Val Glu Val His Thr Ala Gln Thr Gln Pro Arg Glu Glu Gln Phe

275 280 285

5

Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro Ile Met His Gln Asp 290 295 300

10 Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val Asn Ser Ala Ala Phe

305 310 315 320

15 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Arg Pro Lys 325 330 335

Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys Glu Gln Met Ala Lys

20 340 345 350

Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp Phe Phe Pro Glu Asp

355 360 365 25

Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro Ala Glu Asn Tyr Lys 370 375 380

30 Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser Tyr Phe Val Tyr Ser

385 390 395 400

35 Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala Gly Asn Thr Phe Thr 405 410 415

Cys Ser Val Leu His Glu Gly Leu His Asn His His Thr Glu Lys Ser

420 425 430

5

Leu Ser His Ser Pro Gly Lys 435

10 <210> 94

<211> 10 <212> PRT

<213> artificial

15 <220> <223> CDR1 VL 7C4

<400> 94

20 Ser Ala Ser Ser Ser Val Ser Phe Met His

1 5 10

25 <210> 95

<211> 10

<212> PRT <213> artificial

30 <220>

<223> CDR1 VL 7A10/8D9

<400> 95

35 Ser Ala Ser Ser Ser Val Ser Tyr Ile His 1 5 10

<210> 96

<211> 9

5 <212> PRT

<213> artificial

<220>

<223> CDR3 VL L3 10 <400> 96

Gln Gln Trp Thr Ser Asn Pro Pro Phe

1 5

15

<210> 97

<211> 5 <212> PRT

20 <213> artificial

<220> <223> CDR1 VH 7C4

25 <400> 97

Arg Tyr Trp Met His 1 5

30 <210> 98

<211> 17 <212> PRT

<213> artificial

35 <220>

<223> CDR2 VH 5A1

<400> 98

5 Arg Val Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe Lys 1 5 10 15

Thr 10

<210> 99

<211> 17 15 <212> PRT

<213> artificial

<220> <223> CDR2 VH 9G11

20 <400> 99

Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val His Phe Lys

1 5 10 15 25

Thr

30 <210> 100

<211> 17 <212> PRT

<213> artificial

35 <220>

<223> CDR2 VH 9C12

<400> 100

5 Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Ile Lys 1 5 10 15

Thr 10

<210> 101

<211> 12 15 <212> PRT

<213> artificial

<220> <223> CDR3 VH 5A1

20 <400> 101

Leu Gly His Tyr Gly Asn Leu Asn Ala Met Asp Tyr

1 5 10 25

<210> 102 <211> 12

<212> PRT

30 <213> Artificial

<220> <223> CDR3 VH 9D7

35 <400> 102

Leu Gly His Tyr Ser Lys Val Leu Ala Met Asp Tyr

1 5 10

5 <210> 103

<211> 12

<212> PRT <213> artificial

10 <220>

<223> CDR3 VH 7A10/8D9

<400> 103

15 Leu Gly His Tyr Gly Asn Leu Tyr Ala Lys Asp Tyr 1 5 10

<210> 104

20 <211> 106 <212> PRT

<213> artificial

<220> 25 <223> 5A1 VL

<400> 104

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 30 1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

20 25 30

35

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr 35 40 45

5 Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu 10 65 70 75 80

Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr

85 90 95

15

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

20 <210> 105

<211> 118 <212> PRT

<213> artificial 25 <220>

<223> 5A1 VH

<400> 105

30 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Gln Gly Ala

1 5 10 15

35 Ser Vai Lys Vai Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Met His Tyr Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45

5

Gly Arg Val Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe 50 55 60

10 Lys Thr Arg Ala Thr Leu Thr Val Asp Lys Thr Thr Ser Thr Ala Tyr

65 70 75 80

15 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Gly His Tyr Gly Asn Leu Asn Ala Met Asp Tyr Trp Gly

20 100 105 110

Gln Gly Thr Leu Val Thr

115 25

<210> 106 <211> 106

<212> PRT

30 <213> artificial

<220> <223> 9D7 VL

35 <400> 106

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

5 Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr 10 35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

15

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu

65 70 75 80

20 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr

85 90 95

25 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 107 30 <211> 118

<212> PRT

<213> artificial

<220> 35 <223> 9D7 VH

<400> 107

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

5

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

10 Trp Met His Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

15 Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe

50 55 60

Lys Thr Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr

20 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95 25

Ala Arg Leu Gly His Tyr Ser Lys Val Leu Ala Met Asp Tyr Trp Gly 100 105 110

30 Gln Gly Thr Leu Val Thr

115

35 <210> 108

<211> 106

<212> PRT

<213> artificial

<220>

5 <223> 9G11 VL

<400> 108

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

10 1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

20 25 30

15

His Trp Tyr Gln Gln Lys Gln Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

20 Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

25 Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu

65 70 75 80

Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr

30 85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

35

<210> 109

<211> 118 <212> PRT

<213> artificial

5 <220>

<223> 9G11 VH

<400> 109 10 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Met His Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

20 35 40 45

Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val His Phe

50 55 60 25

Lys Thr Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

30 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

35 Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Thr Asp Tyr Trp Gly 100 105 110

Gln Gly Thr Leu Val Thr

115

5

<210> 110 <211> 106

<212> PRT 10 <213> artificial

<220> <223> 7C4 VL

15 <400> 110

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

20 Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Phe Met

20 25 30

25 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

30 50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu

65 70 75 80

35

Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr

85 90 95

5 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 111 10 <211> 118

<212> PRT

<213> artificial

<220> 15 <223> 7C4 VH

<400> 111

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

20 1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Arg Tyr

20 25 30 25

Trp Met His Trp Met Arg Gln Ala Pro Gly Gln Gly Pro Glu Trp Ile 35 40 45

30 Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe

50 55 60

35 Lys Thr Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

5

Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Met Asp Tyr Trp Gly 100 105 110

10 Gln Gly Thr Leu Val Thr

115

15 <210> 112

<211> 106 <212> PRT

<213> artificial

20 <220> <223> L3 VL

<400> 112

25 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

30 20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

35

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

5 Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu 65 70 75 80

Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr 10 85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

15

<210> 113

<211> 118 <212> PRT

20 <213> artificial

<220> <223> L3 VH

25 <400> 113

Gln Val Gln Leu Val Gln Gln Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15

30 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

35 Trp Met His Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45

Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe

50 55 60

5

Lys Thr Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

10 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

15 Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Met Asp Tyr Trp Gly 100 105 110

Gln Gly Thr Leu Val Thr

20 115

<210> 114

<211> 106 25 <212> PRT

<213> artificial

<220>

<223> 7A10 VL

30 <400> 114

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

35

Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Ile

20 25 30

5 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr 35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 10 50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu

65 70 75 80

15

Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Asn

85 90 95

20 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

25 <210> 115

<211> 118

<212> PRT <213> artificial

30 <220>

<223> 7A10 VH

<400> 115

35 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

5

Trp Met His Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45

10 Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe

50 55 60

15 Lys Thr Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

20 85 90 95

Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Lys Asp Tyr Trp Gly

100 105 110 25

Gln Gly Thr Leu Val Thr 115

30 <210> 116

<211> 106 <212> PRT

<213> artificial

35 <220>

<223> 8D9 VL

<400> 116

5 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Ile 10 20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

15

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

20 Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu

65 70 75 80

25 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

30 100 105

<210> 117

<211> 118 35 <212> PRT

<213> artificial

<220> <223> 8D9 VH

5 <400> 117

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15

10 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

15 Trp Met His Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45

Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe

20 50 55 60

Lys Thr Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80 25

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

30 Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Lys Asp Tyr Trp Gly

100 105 110

35 Gln Gly Thr Leu Val Thr 115

<210> 118

<211> 106

5 <212> PRT

<213> artificial

<220>

<223> 9C12 VL 10 <400> 118

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

15

Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

20 25 30

20 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Leu Ile Tyr

35 40 45

25 Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu 30 65 70 75 80

Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr

85 90 95

35

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

5 <210> 119

<211> 118

<212> PRT <213> artificial

10 <220>

<223> 9C12 VH

<400> 119

15 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 20 25 30

Trp Met His Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45 25

Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Ile 50 55 60

30 Lys Thr Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

35 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Met Asp Tyr Trp Gly

100 105 110

5

Gln Gly Thr Leu Val Thr 115

10 <210> 120

<211> 106 <212> PRT

<213> artificial

15 <220> <223> 6B6 VL

<400> 120

20 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

25 Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Leu Ile Tyr

30 35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

35

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu

65 70 75 80

5 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr 85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 10 100 105

<210> 121

<211> 118 15 <212> PRT

<213> artificial

<220> <223> 6B6 VH

20 <400> 121

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15 25

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

30 Trp Met His Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

35 Gly Arg Ile Asp Pro Asn Ser Gly Thr Thr Lys Tyr Asn Val Asn Phe 50 55 60

Lys Thr Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

5

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

10 Ala Arg Leu Gly His Tyr Gly Asn Leu Tyr Ala Met Asp Tyr Trp Gly

100 105 110

15 Gln Gly Thr Leu Val Thr 115

20

Page11 of Page of 64 64

SEQUENCE LISTING SEQUENCE LISTING

<110> H. Lundbeck <110> H. Lundbeck A/S A/S

<120> MonoclonalAnti-Alpha-Synuclein <120> Monoclonal Anti-Alpha-Synuclein Antibodies Antibodies for for Prevent-ing Prevent-ing Tau Tau Aggregation Aggregation <130> <130> 1055 1055

<160> <160> 121 121

<170> <170> PatentInversion PatentIn version3.5 3.5

<210> <210> 1 1 <211> <211> 10 10 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> GM37CDR <223> GM37 CDR11Heavy HeavyChain Chain

<400> <400> 11 Gly Phe Gly Phe Thr Thr Phe Phe Ser Ser Ser Ser Tyr Tyr Ala Ala Met Met Thr Thr 1 1 5 5 10 10

<210> <210> 2 2 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> GM37 CDR2 GM37 CDR2 Heavy Heavy Chain Chain

<400> <400> 2 2

Ala Ile Ala Ile Arg Arg Ser Ser Asn Asn Gly Gly Asp Asp Arg Arg Thr Thr Asp Asp Tyr Tyr Ala Ala Asp Asp Ser Ser Val Val Lys Lys 1 1 5 5 10 10 15 15

Gly Gly

<210> <210> 3 3 <211> <211> 9 9 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> GM37 CDR3 GM37 CDR3 Heavy Heavy Chain Chain

<400> <400> 3 3

Ala Lys Ala Lys Asn Asn Trp Trp Ala Ala Pro Pro Phe Phe Asp Asp Ser Ser 1 1 5 5

<210> <210> 4 4 <211> <211> 11 11 <212> <212> PRT PRT <213> <213> Artificial Artificial

https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H... 14/06/2019 https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H... 14/06/2019

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<220> <220> <223> <223> GM37 CDR1 GM37 CDR1 Light Light Chain Chain

<400> <400> 4 4

Ala Ser Ala Ser Gln Gln Ser Ser Val Val Ser Ser Ser Ser Ser Ser Tyr Tyr Leu Leu Ala Ala 1 1 5 5 10 10

<210> <210> 5 5 <211> <211> 7 7 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> GM37 CDR GM37 CDR 2 2 Light Light Chain Chain

<400> <400> 5 5

Gly Ala Gly Ala Ser SerSer SerArg Arg AlaAla ThrThr 1 1 5 5

<210> <210> 6 6 <211> <211> 9 9 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> GM37 CDR <223> GM37 CDR 33 Light Light Chain Chain

<400> <400> 6 6

Gln Gln Gln Gln Tyr TyrGly GlySer Ser SerSer ProPro Trp Trp Thr Thr 1 1 5 5

<210> <210> 7 7 <211> <211> 113 113 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> GM37 CDR GM37 CDR Heavy Heavy Chain Chain

<400> <400> 7 7

Glu Val Glu Val Gln Gln Leu Leu Leu Leu Glu Glu Ser Ser Gly Gly Gly Gly Gly Gly Leu Leu Val Val Gln Gln Thr Thr Gly Gly Gly Gly 1 1 5 5 10 10 15 15

Ser Leu Ser Leu Arg ArgLeu LeuSer Ser CysCys AlaAla Ala Ala Ser Ser Gly Thr Gly Phe Phe Phe ThrSer PheSer SerTyrSer Tyr 20 20 25 25 30 30

Ala Met Ala Met Thr ThrTrp TrpVal Val ArgArg GlnGln Ala Ala Pro Pro Gly Gly Gly Lys Lys Leu GlyGlu LeuTrp Glu ValTrp Val 35 35 40 40 45 45

Ser Ala Ser Ala Ile IleArg ArgSer Ser AsnAsn GlyGly Asp Asp Arg Arg Thr Tyr Thr Asp Asp Ala TyrAsp AlaSer Asp ValSer Val 50 50 55 55 60 60

https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H... 14/06/2019 https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H...14/06/2019

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Lys Gly Lys Gly Arg ArgPhe PheThr Thr IleIle SerSer Arg Arg Asp Asp Asn Gln Asn Ser Ser Asn GlnThr AsnLeu Thr TyrLeu Tyr 65 65 70 70 75 75 80 80

Leu Gln Leu Gln Met MetAsn AsnSer SerLeuLeu ArgArg Ala Ala Glu Glu Asp Ala Asp Thr Thr Val AlaTyr ValTyr Tyr CysTyr Cys 85 85 90 90 95 95

Ala Lys Ala Lys Asn AsnTrp TrpAla Ala ProPro PhePhe Asp Asp Ser Ser Trp Gln Trp Gly Gly Gly GlnThr GlyLeu Thr ValLeu Val 100 100 105 105 110 110

Thr Thr

<210> <210> 8 8 <211> <211> 108 108 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> GM 37 GM 37 Light LightChain Chain

<400> <400> 8 8

Glu Ile Glu Ile Val ValLeu LeuThr Thr GlnGln SerSer Pro Pro Gly Gly Thr Ser Thr Leu Leu Leu SerSer LeuPro Ser GlyPro Gly 1 1 5 5 10 10 15 15

Glu Arg Glu Arg Ala AlaThr ThrLeu Leu SerSer CysCys Arg Arg Ala Ala Ser Ser Ser Gln Gln Val SerSer ValSer SerSerSer Ser 20 20 25 25 30 30

Tyr Leu Tyr Leu Ala Ala Trp Trp Tyr Tyr Gln Gln Gln Gln Lys Lys Pro Pro Gly Gly Gln Gln Ala Ala Pro Pro Arg Arg Leu Leu Leu Leu 35 35 40 40 45 45

Ile Tyr Gly Ile Tyr GlyAla AlaSer Ser SerSer ArgArg Ala Ala Thr Thr Gly Gly Ile Asp Ile Pro ProArg AspPhe Arg Phe Ser Ser 50 50 55 55 60 60

Gly Ser Gly Ser Gly GlySer SerGly Gly ThrThr AspAsp Phe Phe Thr Thr Leu Ile Leu Thr Thr Ser IleArg SerLeu Arg GluLeu Glu 65 65 70 70 75 75 80 80

Pro Glu Asp Pro Glu AspPhe PheAla AlaValVal TyrTyr Tyr Tyr Cys Cys Gln Gln Gln Gly Gln Tyr TyrSer GlySer Ser ProSer Pro 85 85 90 90 95 95

Trp Thr Trp Thr Phe PheGly GlyGln Gln GlyGly ThrThr Lys Lys Val Val Glu Lys Glu Ile Ile Lys 100 100 105 105

<210> <210> 9 9 <211> <211> 6 6 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> Epitope 112-117 Epitope 112-117

<400> <400> 9 9

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Ile Leu Glu Ile Leu GluAsp AspMet Met ProPro 1 1 5 5

<210> <210> 10 10 <211> <211> 140 140 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> Alpha-synuclein Alpha-synuclein <400> <400> 10 10

Met Asp Met Asp Val ValPhe PheMet Met LysLys GlyGly Leu Leu Ser Ser Lys Lys Lys Ala Ala Glu LysGly GluVal Gly ValVal Val 1 1 5 5 10 10 15 15

Ala Ala Ala Ala Ala AlaGlu GluLys Lys ThrThr LysLys Gln Gln Gly Gly Val Glu Val Ala Ala Ala GluAla AlaGly Ala LysGly Lys 20 20 25 25 30 30

Thr Lys Thr Lys Glu GluGly GlyVal Val LeuLeu TyrTyr Val Val Gly Gly Ser Thr Ser Lys Lys Lys ThrGlu LysGly Glu ValGly Val 35 35 40 40 45 45

Val His Val His Gly GlyVal ValAla Ala ThrThr ValVal Ala Ala Glu Glu Lys Lys Lys Thr Thr Glu LysGln GluVal Gln ThrVal Thr 50 50 55 55 60 60

Asn Val Asn Val Gly GlyGly GlyAla Ala ValVal ValVal Thr Thr Gly Gly Val Ala Val Thr Thr Val AlaAla ValGln Ala LysGln Lys 65 65 70 70 75 75 80 80

Thr Val Thr Val Glu GluGly GlyAla AlaGlyGly SerSer Ile Ile Ala Ala Ala Thr Ala Ala Ala Gly ThrPhe GlyVal Phe LysVal Lys 85 85 90 90 95 95

Lys Asp Lys Asp Gln GlnLeu LeuGly Gly LysLys AsnAsn Glu Glu Glu Glu Gly Pro Gly Ala Ala Gln ProGlu GlnGly Glu IleGly Ile 100 100 105 105 110 110

Leu Glu Leu Glu Asp AspMet MetPro Pro ValVal AspAsp Pro Pro Asp Asp Asn Ala Asn Glu Glu Tyr AlaGlu TyrMet Glu ProMet Pro 115 115 120 120 125 125

Ser Glu Glu Ser Glu GluGly GlyTyr Tyr GlnGln AspAsp Tyr Tyr Glu Glu Pro Pro Glu Ala Glu Ala 130 130 135 135 140 140

<210> <210> 11 11 <211> <211> 165 165 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> A-Syn-AAKK-BAP A-Syn-AAKK-BAP <400> <400> 11 11

https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H... https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H...14/06/201914/06/2019

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Val His Val His Gly Gly Val Val Ala Ala Thr Thr Val Val Ala Ala Glu Glu Lys Lys Thr Thr Lys Lys Glu Glu Gln Gln Val Val Thr Thr 50 50 55 55 60 60

Thr Val Thr Val Glu GluGly GlyAla AlaGlyGly AsnAsn Ile Ile Ala Ala Ala Thr Ala Ala Ala Gly ThrLeu GlyVal Leu LysVal Lys 85 85 90 90 95 95

Lys Asp Lys Asp Gln Gln Leu Leu Ala Ala Lys Lys Gln Gln Asn Asn Glu Glu Glu Glu Gly Gly Phe Phe Leu Leu Gln Gln Glu Glu Gly Gly 100 100 105 105 110 110

Met Val Met Val Asn AsnAsn AsnThr Thr AspAsp IleIle Pro Pro Val Val Asp Glu Asp Pro Pro Asn GluGlu AsnAla Glu TyrAla Tyr 115 115 120 120 125 125

Glu Met Glu Met Pro Pro Pro Pro Glu Glu Glu Glu Glu Glu Tyr Tyr Gln Gln Asp Asp Tyr Tyr Glu Glu Pro Pro Glu Glu Ala Ala Gly Gly 130 130 135 135 140 140

Ser Ala Ser Ala Gly GlyGly GlySer Ser GlyGly GlyGly Leu Leu Asn Asn Asp Phe Asp Ile Ile Glu PheAla GluGln Ala LysGln Lys 145 145 150 150 155 155 160 160

Ile Glu Trp Ile Glu TrpHis HisGlu Glu 165 165

<210> <210> 12 12 <211> <211> 165 165 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> A-Syn-BAAK-BAP A-Syn-BAAK-BAP

<400> <400> 12 12

Met Asp Met Asp Val ValPhe PheMet Met LysLys GlyGly Leu Leu Ser Ser Met Lys Met Ala Ala Glu LysGly GluVal Gly ValVal Val 1 1 5 5 10 10 15 15

Ala Ala Ala Ala Ala AlaGlu GluLys Lys ThrThr LysLys Gln Gln Gly Gly Val Glu Val Thr Thr Ala GluAla AlaGlu Ala LysGlu Lys 20 20 25 25 30 30

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Thr Val Thr Val Glu GluGly GlyAla Ala GlyGly SerSer Ile Ile Ala Ala Ala Thr Ala Ala Ala Gly ThrPhe GlyVal Phe LysVal Lys 85 85 90 90 95 95

Lys Asp Lys Asp Gln GlnLeu LeuAla Ala LysLys GlnGln Asn Asn Glu Glu Glu Phe Glu Gly Gly Leu PheGln LeuGlu Gln GlyGlu Gly 100 100 105 105 110 110

Glu Met Glu Met Pro ProPro ProGlu Glu GluGlu GluGlu Tyr Tyr Gln Gln Asp Glu Asp Tyr Tyr Pro GluGlu ProAla Glu GlyAla Gly 130 130 135 135 140 140

Ser Ala Ser Ala Gly GlyGly GlySer Ser GlyGly GlyGly Leu Leu Asn Asn Asp Asp Ile Glu Ile Phe PheAla GluGln Ala LysGln Lys 145 145 150 150 155 155 160 160

Ile Glu Trp Ile Glu TrpHis HisGlu Glu 165 165

<210> <210> 13 13 <211> <211> 162 162 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> A-Syn-BBAA-BAP A-Syn-BBAA-BAP

<400> <400> 13 13

Thr Lys Thr Lys Glu GluGly GlyVal Val LeuLeu TyrTyr Val Val Gly Gly Ser Thr Ser Lys Lys Arg ThrGlu ArgGly Glu ValGly Val 35 35 40 40 45 45

Val Gln Val Gln Gly GlyVal ValAla Ala SerSer ValVal Ala Ala Glu Glu Lys Lys Lys Thr Thr Glu LysGln GluAla Gln SerAla Ser 50 50 55 55 60 60

His Leu His Leu Gly GlyGly GlyAla Ala ValVal ValVal Thr Thr Gly Gly Val Ala Val Thr Thr Val AlaAla ValGln Ala LysGln Lys 65 65 70 70 75 75 80 80

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Ser Glu Ser Glu Glu GluGly GlyTyr Tyr GlnGln AspAsp Tyr Tyr Glu Glu Pro Ala Pro Glu Glu Gly AlaSer GlyAla Ser GlyAla Gly 130 130 135 135 140 140

Gly Ser Gly Ser Gly GlyGly GlyLeu Leu AsnAsn AspAsp Ile Ile Phe Phe Glu Gln Glu Ala Ala Lys GlnIle LysGlu Ile TrpGlu Trp 145 145 150 150 155 155 160 160

His Glu His Glu

<210> <210> 14 14 <211> <211> 165 165 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> A-Syn-BBKK-BAP A-Syn-BBKK-BAP

<400> <400> 14 14

Ala Ala Ala Ala Ala AlaGlu GluLys Lys ThrThr LysLys Gln Gln Gly Gly Val Glu Val Thr Thr Ala GluAla AlaGlu AlaLysGlu Lys 20 20 25 25 30 30

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Ile Glu Trp Ile Glu TrpHis HisGlu Glu 165 165

<210> <210> 15 15 <211> <211> 141 141 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> A-Syn-120-140_Del-BAP A-Syn-120-140_Del-BAP

<400> <400> 15 15

Ala Ala Ala Ala Ala AlaGlu GluLys Lys ThrThr LysLys Gln Gln Gly Gly Val Glu Val Ala Ala Ala GluAla AlaGly AlaLysGly Lys 20 20 25 25 30 30

Leu Glu Leu Glu Asp AspMet MetPro Pro ValVal AspAsp Gly Gly Ser Ser Ala Gly Ala Gly Gly Ser GlyGly SerGly Gly LeuGly Leu 115 115 120 120 125 125

Asn Asp Asn Asp Ile Ile Phe Phe Glu Glu Ala Ala Gln Gln Lys Lys Ile Ile Glu Glu Trp Trp His His Glu Glu 130 130 135 135 140 140

<210> <210> 16 16 <211> <211> 131 131 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> alpha-synuclein amino <223> alpha-synuclein amino acids acids 1-119 1-119

Page99 of Page of 64 64

<400> <400 > 16 16

Met Ala Met Ala His HisHis HisHis His HisHis HisHis His His Ile Ile Glu Arg Glu Gly Gly Met ArgAsp MetVal Asp PheVal Phe 1 1 5 5 10 10 15 15

Met Lys Met Lys Gly Gly Leu Leu Ser Ser Lys Lys Ala Ala Lys Lys Glu Glu Gly Gly Val Val Val Val Ala Ala Ala Ala Ala Ala Glu Glu 20 20 25 25 30 30

Lys Thr Lys Thr Lys LysGln GlnGly Gly ValVal AlaAla Glu Glu Ala Ala Ala Lys Ala Gly Gly Thr LysLys ThrGlu Lys GlyGlu Gly 35 35 40 40 45 45

Val Leu Val Leu Tyr TyrVal ValGly Gly SerSer LysLys Thr Thr Lys Lys Glu Val Glu Gly Gly Val ValHis ValGly His ValGly Val 50 50 55 55 60 60

Ala Thr Ala Thr Val ValAla AlaGlu Glu LysLys ThrThr Lys Lys Glu Glu Gln Thr Gln Val Val Asn ThrVal AsnGly Val GlyGly Gly 65 65 70 70 75 75 80 80

Ala Val Ala Val Val ValThr ThrGly GlyValVal ThrThr Ala Ala Val Val Ala Lys Ala Gln Gln Thr LysVal ThrGlu Val GlyGlu Gly 85 85 90 90 95 95

Ala Gly Ala Gly Ser Ser Ile Ile Ala Ala Ala Ala Ala Ala Thr Thr Gly Gly Phe Phe Val Val Lys Lys Lys Lys Asp Asp Gln Gln Leu Leu 100 100 105 105 110 110

Gly Lys Gly Lys Asn AsnGlu GluGlu Glu GlyGly AlaAla Pro Pro Gln Gln Glu Ile Glu Gly Gly Leu IleGlu LeuAsp Glu MetAsp Met 115 115 120 120 125 125

Pro Val Pro Val Asp Asp 130 130

<210> <210> 17 17 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> kappa (LC kappa (LCconstant constant region) region)

<400> <400> 17 17

Thr Val Thr Val Ala Ala Ala Ala Pro Pro Ser Ser Val Val Phe Phe Ile Ile Phe Phe Pro Pro Pro Pro Ser Ser Asp Asp Glu Glu Gln Gln 1 1 5 5 10 10 15 15

Leu Lys Leu Lys Ser SerGly GlyThr Thr AlaAla SerSer Val Val Val Val Cys Leu Cys Leu Leu Asn LeuAsn AsnPhe AsnTyrPhe Tyr 20 20 25 25 30 30

Pro Arg Pro Arg Glu GluAla AlaLys Lys ValVal GlnGln Trp Trp Lys Lys Val Asn Val Asp Asp Ala AsnLeu AlaGln Leu SerGln Ser 35 35 40 40 45 45

Gly Asn Gly Asn Ser SerGln GlnGlu Glu SerSer ValVal Thr Thr Glu Glu Gln Ser Gln Asp Asp Lys SerAsp LysSer Asp ThrSer Thr 50 50 55 55 60 60

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Tyr Ser Tyr Ser Leu LeuSer SerSer Ser ThrThr LeuLeu Thr Thr Leu Leu Ser Ala Ser Lys Lys Asp AlaTyr AspGlu Tyr LysGlu Lys 65 65 70 70 75 75 80 80

His Lys His Lys Val ValTyr TyrAla AlaCysCys GluGlu Val Val Thr Thr His Gly His Gln Gln Leu GlySer LeuSer Ser ProSer Pro 85 85 90 90 95 95

Val Thr Val Thr Lys LysSer SerPhe Phe AsnAsn ArgArg Gly Gly Glu Glu Cys Cys 100 100 105 105

<210> <210> 18 18 <211> <211> 329 329 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> IgG1 (HC Constant IgG1 (HC Constantregion) region)

<400> <400> 18 18

Ala Ser Ala Ser Thr ThrLys LysGly Gly ProPro SerSer Val Val Phe Phe Pro Ala Pro Leu Leu Pro AlaSer ProSer Ser LysSer Lys 1 1 5 5 10 10 15 15

Ser Thr Ser Thr Ser SerGly GlyGly Gly ThrThr AlaAla Ala Ala Leu Leu Gly Leu Gly Cys Cys Val LeuLys ValAsp Lys TyrAsp Tyr 20 20 25 25 30 30

Phe Pro Phe Pro Glu GluPro ProVal Val ThrThr ValVal Ser Ser Trp Trp Asn Gly Asn Ser Ser Ala GlyLeu AlaThr Leu SerThr Ser 35 35 40 40 45 45

Gly Val Gly Val His HisThr ThrPhe Phe ProPro AlaAla Val Val Leu Leu Gln Ser Gln Ser Ser Gly SerLeu GlyTyr Leu SerTyr Ser 50 50 55 55 60 60

Leu Ser Leu Ser Ser SerVal ValVal Val ThrThr ValVal Pro Pro Ser Ser Ser Leu Ser Ser Ser Gly LeuThr GlyGln Thr ThrGln Thr 65 65 70 70 75 75 80 80

Tyr Ile Tyr Ile Cys CysAsn AsnVal ValAsnAsn HisHis Lys Lys Pro Pro Ser Thr Ser Asn Asn Lys ThrVal LysAsp Val LysAsp Lys 85 85 90 90 95 95

Arg Val Arg Val Glu GluPro ProLys Lys SerSer CysCys Asp Asp Lys Lys Thr Thr Thr His His Cys ThrPro CysPro Pro CysPro Cys 100 100 105 105 110 110

Pro Ala Pro Ala Pro ProGlu GluLeu Leu LeuLeu GlyGly Gly Gly Pro Pro Ser Phe Ser Val Val Leu PhePhe LeuPro Phe ProPro Pro 115 115 120 120 125 125

Lys Pro Lys Pro Lys LysAsp AspThr Thr LeuLeu MetMet Ile Ile Ser Ser Arg Pro Arg Thr Thr Glu ProVal GluThr Val CysThr Cys 130 130 135 135 140 140

Val Val Val Val Val Val Asp Asp Val Val Ser Ser His His Glu Glu Asp Asp Pro Pro Glu Glu Val Val Lys Lys Phe Phe Asn Asn Trp Trp 145 145 150 150 155 155 160 160

Tyr Val Tyr Val Asp AspGly GlyVal Val GluGlu ValVal His His Asn Asn Ala Thr Ala Lys Lys Lys ThrPro LysArg Pro GluArg Glu 165 165 170 170 175 175

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Glu Gln Glu Gln Tyr Tyr Asn Asn Ser Ser Thr Thr Tyr Tyr Arg Arg Val Val Val Val Ser Ser Val Val Leu Leu Thr Thr Val Val Leu Leu 180 180 185 185 190 190

His Gln His Gln Asp Asp Trp Trp Leu Leu Asn Asn Gly Gly Lys Lys Glu Glu Tyr Tyr Lys Lys Cys Cys Lys Lys Val Val Ser Ser Asn Asn 195 195 200 200 205 205

Lys Ala Lys Ala Leu Leu Pro Pro Ala Ala Pro Pro Ile Ile Glu Glu Lys Lys Thr Thr Ile Ile Ser Ser Lys Lys Ala Ala Lys Lys Gly Gly 210 210 215 215 220 220

Gln Pro Gln Pro Arg Arg Glu Glu Pro Pro Gln Gln Val Val Tyr Tyr Thr Thr Leu Leu Pro Pro Pro Pro Ser Ser Arg Arg Glu Glu Glu Glu 225 225 230 230 235 235 240 240

Met Thr Met Thr Lys LysAsn AsnGln Gln ValVal SerSer Leu Leu Thr Thr Cys Val Cys Leu Leu Lys ValGly LysPhe Gly TyrPhe Tyr 245 245 250 250 255 255

Pro Ser Pro Ser Asp AspIle IleAla Ala ValVal GluGlu Trp Trp Glu Glu Ser Ser Asn Gln Asn Gly GlyPro GlnGlu Pro AsnGlu Asn 260 260 265 265 270 270

Asn Tyr Asn Tyr Lys LysThr ThrThr Thr ProPro ProPro Val Val Leu Leu Asp Asp Asp Ser Ser Gly AspSer GlyPhe Ser PhePhe Phe 275 275 280 280 285 285

Leu Tyr Leu Tyr Ser SerLys LysLeu Leu ThrThr ValVal Asp Asp Lys Lys Ser Trp Ser Arg Arg Gln TrpGln GlnGly Gln AsnGly Asn 290 290 295 295 300 300

Val Phe Val Phe Ser SerCys CysSer Ser ValVal MetMet His His Glu Glu Ala His Ala Leu Leu Asn HisHis AsnTyr His ThrTyr Thr 305 305 310 310 315 315 320 320

Gln Lys Gln Lys Ser Ser Leu Leu Ser Ser Leu Leu Ser Ser Pro Pro Gly Gly 325 325

<210> <210> 19 19 <211> <211> 4 4 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> GM285 epitope112-115 GM285 epitope 112-115

<400> <400> 19 19

Ile Leu Glu Ile Leu GluAsp Asp 1 1

<210> <210> 20 20 <211> <211> 13 13 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> GM285 CDR1 <223> GM285 CDR1 Heavy Heavy Chain Chain

Page 12 Page 12of of 64 64

<400> <400> 20 20

Ala Ala Ala Ala Ser Ser Gly Gly Phe Phe Thr Thr Phe Phe Ser Ser Arg Arg Phe Phe Thr Thr Met Met Thr Thr 1 1 5 5 10 10

<210> <210> 21 21 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> GM285 CDR2 GM285 CDR2 Heavy Heavy Chain Chain

<400> <400> 21 21

Ala Ile Ala Ile Ser Ser Gly Gly Ser Ser Gly Gly Gly Gly Gly Gly Thr Thr Ser Ser Tyr Tyr Ala Ala Asp Asp Ser Ser Val Val Lys Lys 1 1 5 5 10 10 15 15

Gly Gly

<210> <210> 22 22 <211> <211> 9 9 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> GM285 CDR3 <223> GM285 CDR3 Heavy Heavy Chain Chain

<400> <400> 22 22

Ala Lys Ala Lys Asn Asn Trp Trp Ala Ala Pro Pro Phe Phe Asp Asp Tyr Tyr 1 1 5 5

<210> <210> 23 23 <211> <211> 12 12 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> GM285 CDR1 GM285 CDR1 Light Light Chain Chain

<400> <400> 23 23

Arg Ala Arg Ala Ser Ser Gln Gln Ser Ser Val Val Ser Ser Arg Arg Ser Ser Tyr Tyr Leu Leu Ala Ala 1 1 5 5 10 10

<210> <210> 24 24 <211> <211> 7 7 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> GM285 CDR2 GM285 CDR2 Light Light Chain Chain

<400> <400> 24 24

Gly Ala Gly Ala Ser Ser Ser Ser Arg Arg Ala Ala Thr Thr 1 1 5 5

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<210> <210> 25 25 <211> <211> 9 9 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> GM285 CDR3 GM285 CDR3Light LightChain Chain

<400> <400> 25 25

Gln Gln Gln Gln Tyr TyrGly GlySer Ser SerSer ProPro Trp Trp Thr Thr 1 1 5 5

<210> <210> 26 26 <211> <211> 113 113 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> GM285 VH GM285 VH

<400> <400> 26 26

Glu Val Glu Val Gln GlnLeu LeuLeu Leu GluGlu SerSer Gly Gly Gly Gly Gly Val Gly Leu Leu Gln ValPro GlnGly Pro GlyGly Gly 1 1 5 5 10 10 15 15

Ser Leu Arg Ser Leu ArgLeu LeuSer Ser CysCys AlaAla Ala Ala Ser Ser Gly Gly Phe Phe Phe Thr ThrSer PheArg SerPheArg Phe 20 20 25 25 30 30

Thr Met Thr Met Thr ThrTrp TrpVal Val ArgArg GlnGln Ala Ala Pro Pro Gly Gly Gly Lys Lys Leu GlyGlu LeuTrp Glu ValTrp Val 35 35 40 40 45 45

Ser Ala Ile Ser Ala IleSer SerGly Gly SerSer GlyGly Gly Gly Gly Gly Thr Thr Ser Ala Ser Tyr TyrAsp AlaSer Asp ValSer Val 50 50 55 55 60 60

Lys Gly Lys Gly Arg ArgLeu LeuThr Thr ValVal SerSer Arg Arg Asp Asp Asn Lys Asn Ser Ser Asn LysThr AsnLeu Thr TyrLeu Tyr 65 65 70 70 75 75 80 80

Ala Lys Ala Lys Asn AsnTrp TrpAla Ala ProPro PhePhe Asp Asp Tyr Tyr Trp Gln Trp Gly Gly Gly GlnThr GlyLeu Thr ValLeu Val 100 100 105 105 110 110

Thr Thr

<210> <210> 27 27 <211> <211> 108 108 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220>

Page 14 Page 14of of 64 64

<223> 223> GM285 VL GM285 VL

<400> 400> 27 27

Glu Arg Glu Arg Ala AlaThr ThrLeu Leu SerSer CysCys Arg Arg Ala Ala Ser Ser Ser Gln Gln Val SerSer ValArg SerSerArg Ser 20 20 25 25 30 30

Gly Ser Gly Ser Gly Gly Ser Ser Gly Gly Thr Thr Asp Asp Phe Phe Thr Thr Leu Leu Thr Thr Val Val Ser Ser Arg Arg Leu Leu Glu Glu 65 65 70 70 75 75 80 80

Pro Glu Pro Glu Asp AspPhe PheAla AlaValVal TyrTyr Tyr Tyr Cys Cys Gln Tyr Gln Gln Gln Gly TyrSer GlySer Ser ProSer Pro 85 85 90 90 95 95

<210> <210> 28 28 <211> <211> 329 329 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> GM285 IgG1 GM285 IgG1 constant constant region region

<400> <400> 28 28

Ser Thr Ser Ser Thr SerGly GlyGly Gly ThrThr AlaAla Ala Ala Leu Leu Gly Gly Cys Val Cys Leu LeuLys ValAsp LysTyrAsp Tyr 20 20 25 25 30 30

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Val Val Val Val Val ValAsp AspVal Val SerSer HisHis Glu Glu Asp Asp Pro Val Pro Glu Glu Lys ValPhe LysAsn Phe TrpAsn Trp 145 145 150 150 155 155 160 160

Glu Gln Glu Gln Tyr TyrAsn AsnSer Ser ThrThr TyrTyr Arg Arg Val Val Val Val Val Ser Ser Leu ValThr LeuVal Thr LeuVal Leu 180 180 185 185 190 190

Gln Pro Gln Pro Arg ArgGlu GluPro Pro GlnGln ValVal Tyr Tyr Thr Thr Leu Pro Leu Pro Pro Ser ProArg SerGlu Arg GluGlu Glu 225 225 230 230 235 235 240 240

Pro Ser Pro Ser Asp AspIle IleAla Ala ValVal GluGlu Trp Trp Glu Glu Ser Gly Ser Asn Asn Gln GlyPro GlnGlu Pro AsnGlu Asn 260 260 265 265 270 270

Gln Lys Gln Lys Ser Ser Leu Leu Ser Ser Leu Leu Ser Ser Pro Pro Gly Gly 325 325

<210> <210> 29 29 <211> <211> 106 106

Page16 Page 16of of 64 64

<212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> GM285 Kappa GM285 Kappachain chain

<400> <400> 29 29

Thr Val Thr Val Ala AlaAla AlaPro Pro SerSer ValVal Phe Phe Ile Ile Phe Pro Phe Pro Pro Ser ProAsp SerGlu Asp GlnGlu Gln 1 1 5 5 10 10 15 15

Leu Lys Leu Lys Ser SerGly GlyThr Thr AlaAla SerSer Val Val Val Val Cys Leu Cys Leu Leu Asn LeuAsn AsnPhe Asn TyrPhe Tyr 20 20 25 25 30 30

His Lys His Lys Val ValTyr TyrAla Ala CysCys GluGlu Val Val Thr Thr His Gly His Gln Gln Leu GlySer LeuSer Ser ProSer Pro 85 85 90 90 95 95

<210> <210> 30 30 <211> <211> 113 113 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> GM37 Variant GM37 Variant 1 1 heavy heavy chain chain

<400> <400> 30 30

Glu Val Glu Val Gln GlnLeu LeuLeu Leu GluGlu SerSer Gly Gly Gly Gly Gly Val Gly Leu Leu Gln ValThr GlnGly Thr GlyGly Gly 1 1 5 5 10 10 15 15

Ser Leu Arg Ser Leu ArgLeu LeuSer Ser CysCys AlaAla Ala Ala Ser Ser Gly Gly Phe Phe Phe Thr ThrSer PheSer SerTyrSer Tyr 20 20 25 25 30 30

Ser Ala Ile Ser Ala IleArg ArgSer Ser SerSer GlyGly Asp Asp Arg Arg Thr Thr Asp Ala Asp Tyr TyrAsp AlaSer Asp ValSer Val 50 50 55 55 60 60

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Thr Thr

<210> <210> 31 31 <211> <211> 113 113 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> GM 37 GM 37 variant variant2 2heavy heavy chain chain

<400> <400> 31 31

Ser Leu Ser Leu Arg ArgLeu LeuSer Ser CysCys AlaAla Ala Ala Ser Ser Gly Gly Phe Phe Phe Thr ThrSer PheSer Ser TyrSer Tyr 20 20 25 25 30 30

Ser Ala Ser Ala Ile IleArg ArgSer Ser GlnGln GlyGly Asp Asp Arg Arg Thr Tyr Thr Asp Asp Ala TyrAsp AlaSer Asp ValSer Val 50 50 55 55 60 60

Leu Gln Leu Gln Met MetAsn AsnSer Ser LeuLeu ArgArg Ala Ala Glu Glu Asp Ala Asp Thr Thr Val AlaTyr ValTyr Tyr CysTyr Cys 85 85 90 90 95 95

Thr Thr

<210> <210> 32 32 <211> <211> 113 113 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> GM 37 <223> GM 37 variant variant 33 heavy heavy chain chain

Page 18 Page 18of of 64 64

<400> 400> 32 32

Ser Ala Ile Ser Ala IleArg ArgSer Ser HisHis GlyGly Asp Asp Arg Arg Thr Thr Asp Ala Asp Tyr TyrAsp AlaSer Asp ValSer Val 50 50 55 55 60 60

Thr Thr

<210> <210> 33 33 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> GM37 variant GM37 variant1 1heavy heavy chain chain CDR CDR 2 2

<400> <400> 33 33

Ala Ile Ala Ile Arg ArgSer SerSer Ser GlyGly AspAsp Arg Arg Thr Thr Asp Ala Asp Tyr Tyr Asp AlaSer AspVal Ser LysVal Lys 1 1 5 5 10 10 15 15

Gly Gly

<210> <210> 34 34 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> GM37 variant GM37 variant 2 2 CDR CDR 2 2 heavy heavy chain chain

<400> <400> 34 34

Ala Ile Ala Ile Arg ArgSer SerGln Gln GlyGly AspAsp Arg Arg Thr Thr Asp Ala Asp Tyr Tyr Asp AlaSer AspVal Ser LysVal Lys 1 1 5 5 10 10 15 15

https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H... ttps://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H...14/06/201914/06/2019

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Gly Gly

<210> <210> 35 35 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> GM37 variant <223> GM37 variant 33 CDR CDR 22 heavy heavy chain chain

<400> <400> 35 35

Ala Ile Ala Ile Arg ArgSer SerHis His GlyGly AspAsp Arg Arg Thr Thr Asp Ala Asp Tyr Tyr Asp AlaSer AspVal Ser LysVal Lys 1 1 5 5 10 10 15 15

Gly Gly

<210> <210> 36 36 <211> <211> 5 5 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> 9E4 bindingepitope 9E4 binding epitope

<400> <400> 36 36

Asn Glu Asn Glu Ala Ala Tyr Tyr Glu Glu 1 1 5 5

<210> <210> 37 37 <211> <211> 134 134 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> HUMAN Beta-synuclein HUMAN Beta-synuclein

<400> <400> 37 37

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His Leu His Leu Gly GlyGly GlyAla Ala ValVal PhePhe Ser Ser Gly Gly Ala Asn Ala Gly Gly Ile AsnAla IleAla Ala AlaAla Ala 65 65 70 70 75 75 80 80

Thr Gly Thr Gly Leu Leu Val Val Lys Lys Arg Arg Glu Glu Glu Glu Phe Phe Pro Pro Thr Thr Asp Asp Leu Leu Lys Lys Pro Pro Glu Glu 85 85 90 90 95 95

Glu Val Glu Val Ala AlaGln GlnGlu Glu AlaAla AlaAla Glu Glu Glu Glu Pro Ile Pro Leu Leu Glu IlePro GluLeu Pro MetLeu Met 100 100 105 105 110 110

Glu Pro Glu Pro Glu Glu Gly Gly Glu Glu Ser Ser Tyr Tyr Glu Glu Asp Asp Pro Pro Pro Pro Gln Gln Glu Glu Glu Glu Tyr Tyr Gln Gln 115 115 120 120 125 125

Glu Tyr Glu Tyr Glu GluPro ProGlu Glu AlaAla 130 130

<210> <210> 38 38 <211> <211> 127 127 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> HUMAN Gamma-synuclein HUMAN Gamma-synuclein

<400> <400> 38 38

Met Asp Met Asp Val ValPhe PheLys Lys LysLys GlyGly Phe Phe Ser Ser Ile Lys Ile Ala Ala Glu LysGly GluVal Gly ValVal Val 1 1 5 5 10 10 15 15

Gly Ala Gly Ala Val ValGlu GluLys Lys ThrThr LysLys Gln Gln Gly Gly Val Glu Val Thr Thr Ala GluAla AlaGlu AlaLysGlu Lys 20 20 25 25 30 30

Thr Lys Thr Lys Glu GluGly GlyVal Val MetMet TyrTyr Val Val Gly Gly Ala Thr Ala Lys Lys Lys ThrGlu LysAsn Glu ValAsn Val 35 35 40 40 45 45

Val Gln Val Gln Ser Ser Val Val Thr Thr Ser Ser Val Val Ala Ala Glu Glu Lys Lys Thr Thr Lys Lys Glu Glu Gln Gln Ala Ala Asn Asn 50 50 55 55 60 60

Ala Val Ala Val Ser SerGlu GluAla Ala ValVal ValVal Ser Ser Ser Ser Val Thr Val Asn Asn Val ThrAla ValThr Ala LysThr Lys 65 65 70 70 75 75 80 80

Thr Val Thr Val Glu Glu Glu Glu Ala Ala Glu Glu Asn Asn Ile Ile Ala Ala Val Val Thr Thr Ser Ser Gly Gly Val Val Val Val Arg Arg 85 85 90 90 95 95

Lys Glu Lys Glu Asp AspLeu LeuArg Arg ProPro SerSer Ala Ala Pro Pro Gln Glu Gln Gln Gln Gly GluGlu GlyAla Glu SerAla Ser 100 100 105 105 110 110

Lys Glu Lys Glu Lys Lys Glu Glu Glu Glu Val Val Ala Ala Glu Glu Glu Glu Ala Ala Gln Gln Ser Ser Gly Gly Gly Gly Asp Asp 115 115 120 120 125 125

<210> <210> 39 39 <211> <211> 140 140

Page21 Page 21of of 64 64

<212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> alpha-synuclein ortholog alpha-synuclein ortholog forfor Cynomolgus Cynomolgus monkey monkey

<400> <400> 39 39

Thr Val Thr Val Glu GluGly GlyAla Ala GlyGly SerSer Ile Ile Ala Ala Ala Thr Ala Ala Ala Gly ThrPhe GlyIle Phe LysIle Lys 85 85 90 90 95 95

Leu Gln Leu Gln Asp AspMet MetPro Pro ValVal AspAsp Pro Pro Asp Asp Asn Ala Asn Glu Glu Tyr AlaGlu TyrMet Glu ProMet Pro 115 115 120 120 125 125

Ser Glu Ser Glu Glu GluGly GlyTyr Tyr GlnGln AspAsp Tyr Tyr Glu Glu Pro Pro Glu Ala Glu Ala 130 130 135 135 140 140

<210> <210> 40 40 <211> <211> 140 140 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> alpha-synucleinortholog alpha-synuclein ortholog forfor Rat Rat

<400> <400> 40 40

https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H... 14/06/2019 hhttps://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H...14/06/2019

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Val His Gly Val Thr Thr Val Ala Glu Lys Thr Lys Glu Gln Val Thr 50 50 55 55 60 60

Thr Val Thr Val Glu GluGly GlyAla AlaGlyGly AsnAsn Ile Ile Ala Ala Ala Thr Ala Ala Ala Gly ThrPhe GlyVal Phe LysVal Lys 85 85 90 90 95 95

Lys Asp Lys Asp Gln GlnMet MetGly Gly LysLys GlyGly Glu Glu Glu Glu Gly Pro Gly Tyr Tyr Gln ProGlu GlnGly Glu IleGly Ile 100 100 105 105 110 110

Leu Glu Leu Glu Asp AspMet MetPro Pro ValVal AspAsp Pro Pro Ser Ser Ser Ala Ser Glu Glu Tyr AlaGlu TyrMet Glu ProMet Pro 115 115 120 120 125 125

Ser Glu Ser Glu Glu GluGly GlyTyr Tyr GlnGln AspAsp Tyr Tyr Glu Glu Pro Ala Pro Glu Glu Ala 130 130 135 135 140 140

<210> <210> 41 41 <211> <211> 140 140 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> alpha-synucleinortholog alpha-synuclein ortholog forfor Mouse Mouse

<400> <400> 41 41

Val His Val His Gly Gly Val Val Thr Thr Thr Thr Val Val Ala Ala Glu Glu Lys Lys Thr Thr Lys Lys Glu Glu Gln Gln Val Val Thr Thr 50 50 55 55 60 60

Page 23 Page 23of of 64 64

Leu Glu Leu Glu Asp AspMet MetPro Pro ValVal AspAsp Pro Pro Gly Gly Ser Ala Ser Glu Glu Tyr AlaGlu TyrMet Glu ProMet Pro 115 115 120 120 125 125

<210> <210> 42 42 <211> <211> 446 446 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9E4 HC 9E4 HC

<400> <400> 42 42

Glu Val Glu Val Gln GlnLeu LeuVal Val GluGlu SerSer Gly Gly Gly Gly Gly Val Gly Leu Leu Gln ValPro GlnGly Pro GlyGly Gly 1 1 5 5 10 10 15 15

Ser Leu Ser Leu Arg ArgLeu LeuSer Ser CysCys AlaAla Ala Ala Ser Ser Gly Thr Gly Phe Phe Phe ThrSer PheAsn Ser TyrAsn Tyr 20 20 25 25 30 30

Gly Met Gly Met Ser SerTrp TrpVal Val ArgArg GlnGln Ala Ala Pro Pro Gly Gly Gly Lys Lys Leu GlyGlu LeuTrp Glu ValTrp Val 35 35 40 40 45 45

Ala Ser Ala Ser Ile IleSer SerSer Ser GlyGly GlyGly Gly Gly Ser Ser Thr Tyr Thr Tyr Tyr Pro TyrAsp ProAsn Asp ValAsn Val 50 50 55 55 60 60

Lys Gly Lys Gly Arg ArgPhe PheThr Thr IleIle SerSer Arg Arg Asp Asp Asn Lys Asn Ala Ala Asn LysSer AsnLeu Ser TyrLeu Tyr 65 65 70 70 75 75 80 80

Ala Arg Ala Arg Gly GlyGly GlyAla Ala GlyGly IleIle Asp Asp Tyr Tyr Trp Gln Trp Gly Gly Gly GlnThr GlyLeu Thr ValLeu Val 100 100 105 105 110 110

Thr Val Thr Val Ser SerSer SerAla Ala SerSer ThrThr Lys Lys Gly Gly Pro Val Pro Ser Ser Phe ValPro PheLeu Pro AlaLeu Ala 115 115 120 120 125 125

Pro Ser Pro Ser Ser SerLys LysSer Ser ThrThr SerSer Gly Gly Gly Gly Thr Ala Thr Ala Ala Leu AlaGly LeuCys Gly LeuCys Leu 130 130 135 135 140 140

Val Lys Val Lys Asp Asp Tyr Tyr Phe Phe Pro Pro Glu Glu Pro Pro Val Val Thr Thr Val Val Ser Ser Trp Trp Asn Asn Ser Ser Gly Gly 145 145 150 150 155 155 160 160

Ala Leu Ala Leu Thr ThrSer SerGly Gly ValVal HisHis Thr Thr Phe Phe Pro Val Pro Ala Ala Leu ValGln LeuSer Gln SerSer Ser 165 165 170 170 175 175

Gly Leu Gly Leu Tyr Tyr Ser Ser Leu Leu Ser Ser Ser Ser Val Val Val Val Thr Thr Val Val Pro Pro Ser Ser Ser Ser Ser Ser Leu Leu 180 180 185 185 190 190

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Gly Thr Gly Thr Gln Gln Thr Thr Tyr Tyr Ile Ile Cys Cys Asn Asn Val Val Asn Asn His His Lys Lys Pro Pro Ser Ser Asn Asn Thr Thr 195 195 200 200 205 205

Lys Val Lys Val Asp AspLys LysArg Arg ValVal GluGlu Pro Pro Lys Lys Ser Asp Ser Cys Cys Lys AspThr LysHis Thr ThrHis Thr 210 210 215 215 220 220

Cys Pro Cys Pro Pro ProCys CysPro Pro AlaAla ProPro Glu Glu Leu Leu Leu Gly Leu Gly Gly Pro GlySer ProVal Ser PheVal Phe 225 225 230 230 235 235 240 240

Leu Phe Leu Phe Pro ProPro ProLys Lys ProPro LysLys Asp Asp Thr Thr Leu Ile Leu Met Met Ser IleArg SerThr Arg ProThr Pro 245 245 250 250 255 255

Glu Val Glu Val Thr ThrCys CysVal Val ValVal ValVal Asp Asp Val Val Ser Glu Ser His His Asp GluPro AspGlu Pro ValGlu Val 260 260 265 265 270 270

Lys Phe Lys Phe Asn AsnTrp TrpTyr Tyr ValVal AspAsp Gly Gly Val Val Glu His Glu Val Val Asn HisAla AsnLys Ala ThrLys Thr 275 275 280 280 285 285

Lys Pro Lys Pro Arg ArgGlu GluGlu Glu GlnGln TyrTyr Asn Asn Ser Ser Thr Arg Thr Tyr Tyr Val ArgVal ValSer Val ValSer Val 290 290 295 295 300 300

Leu Thr Leu Thr Val ValLeu LeuHis His GlnGln AspAsp Trp Trp Leu Leu Asn Lys Asn Gly Gly Glu LysTyr GluLys Tyr CysLys Cys 305 305 310 310 315 315 320 320

Lys Val Lys Val Ser SerAsn AsnLys Lys AlaAla LeuLeu Pro Pro Ala Ala Pro Glu Pro Ile Ile Lys GluThr LysIle Thr SerIle Ser 325 325 330 330 335 335

Lys Ala Lys Ala Lys LysGly GlyGln Gln ProPro ArgArg Glu Glu Pro Pro Gln Tyr Gln Val Val Thr TyrLeu ThrPro Leu ProPro Pro 340 340 345 345 350 350

Ser Arg Ser Arg Glu GluGlu GluMet Met ThrThr LysLys Asn Asn Gln Gln Val Leu Val Ser Ser Thr LeuCys ThrLeu Cys ValLeu Val 355 355 360 360 365 365

Lys Gly Lys Gly Phe PheTyr TyrPro Pro SerSer AspAsp Ile Ile Ala Ala Val Trp Val Glu Glu Glu TrpSer GluAsn Ser GlyAsn Gly 370 370 375 375 380 380

Gln Pro Gln Pro Glu Glu Asn Asn Asn Asn Tyr Tyr Lys Lys Thr Thr Thr Thr Pro Pro Pro Pro Val Val Leu Leu Asp Asp Ser Ser Asp Asp 385 385 390 390 395 395 400 400

Gly Ser Gly Ser Phe PhePhe PheLeu Leu TyrTyr SerSer Lys Lys Leu Leu Thr Asp Thr Val Val Lys AspSer LysArg Ser TrpArg Trp 405 405 410 410 415 415

Gln Gln Gln Gln Gly GlyAsn AsnVal Val PhePhe SerSer Cys Cys Ser Ser Val His Val Met Met Glu HisAla GluLeu Ala HisLeu His 420 420 425 425 430 430

Asn His Asn His Tyr TyrThr ThrGln Gln LysLys SerSer Leu Leu Ser Ser Leu Pro Leu Ser Ser Gly ProLys Gly Lys 435 435 440 440 445 445

Page 25 Page 25of of 64 64

<210> <210> 43 43 <211> <211> 220 220 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9E4 LC 9E4 LC

<400> <400> 43 43

Asp Ile Asp Ile Gln Gln Met Met Thr Thr Gln Gln Ser Ser Pro Pro Ser Ser Ser Ser Leu Leu Ser Ser Ala Ala Ser Ser Val Val Gly Gly 1 1 5 5 10 10 15 15

Asp Arg Asp Arg Val ValThr ThrIle Ile ThrThr CysCys Lys Lys Ser Ser Ile Thr Ile Gln Gln Leu ThrLeu LeuTyr LeuSerTyr Ser 20 20 25 25 30 30

Ser Asn Gln Ser Asn GlnLys LysAsn Asn TyrTyr LeuLeu Ala Ala Trp Trp Phe Phe Gln Lys Gln Gln GlnPro LysGly Pro LysGly Lys 35 35 40 40 45 45

Ala Pro Ala Pro Lys LysLeu LeuLeu Leu IleIle TyrTyr Trp Trp Ala Ala Ser Arg Ser Ile Ile Lys ArgSer LysGly Ser ValGly Val 50 50 55 55 60 60

Pro Ser Pro Ser Arg ArgPhe PheSer Ser GlyGly SerSer Gly Gly Ser Ser Gly Gly Thr Phe Thr Asp AspThr PheLeu Thr ThrLeu Thr 65 65 70 70 75 75 80 80

Ile Ser Ser Ile Ser SerLeu LeuGln GlnProPro GluGlu Asp Asp Leu Leu Ala Ala Thr Tyr Thr Tyr TyrCys TyrGln Cys Gln Gln Gln 85 85 90 90 95 95

Tyr Tyr Tyr Tyr Ser SerTyr TyrPro Pro LeuLeu ThrThr Phe Phe Gly Gly Gly Thr Gly Gly Gly Lys ThrLeu LysGlu Leu IleGlu Ile 100 100 105 105 110 110

Lys Arg Lys Arg Thr ThrVal ValAla Ala AlaAla ProPro Ser Ser Val Val Phe Phe Phe Ile Ile Pro PhePro ProSer Pro AspSer Asp 115 115 120 120 125 125

Glu Gln Glu Gln Leu LeuLys LysSer Ser GlyGly ThrThr Ala Ala Ser Ser Val Cys Val Val Val Leu CysLeu LeuAsn Leu AsnAsn Asn 130 130 135 135 140 140

Phe Tyr Phe Tyr Pro ProArg ArgGlu Glu AlaAla LysLys Val Val Gln Gln Trp Trp Lys Asp Lys Val ValAsn AspAla Asn LeuAla Leu 145 145 150 150 155 155 160 160

Gln Ser Gln Ser Gly GlyAsn AsnSer Ser GlnGln GluGlu Ser Ser Val Val Thr Gln Thr Glu Glu Asp GlnSer AspLys Ser AspLys Asp 165 165 170 170 175 175

Ser Thr Tyr Ser Thr TyrSer SerLeu Leu SerSer SerSer Thr Thr Leu Leu Thr Thr Leu Lys Leu Ser SerAla LysAsp Ala TyrAsp Tyr 180 180 185 185 190 190

Glu Lys Glu Lys His HisLys LysVal Val TyrTyr AlaAla Cys Cys Glu Glu Val His Val Thr Thr Gln HisGly GlnLeu Gly SerLeu Ser 195 195 200 200 205 205

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Ser Pro Ser Pro Val ValThr ThrLys Lys SerSer PhePhe Asn Asn Arg Arg Gly Cys Gly Glu Glu Cys 210 210 215 215 220 220

<210> <210> 44 44 <211> <211> 10 10 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9E4 CDR1 Heavy 9E4 CDR1 HeavyChain Chain

<400> <400> 44 44

Gly Phe Gly Phe Thr ThrPhe PheSer Ser AsnAsn TyrTyr Gly Gly Met Met Ser Ser 1 1 5 5 10 10

<210> <210> 45 45 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9E4 CDR2 Heavy 9E4 CDR2 HeavyChain Chain

<400> <400> 45 45

Ser Ile Ser Ile Ser SerSer SerGly Gly GlyGly GlyGly Ser Ser Thr Thr Tyr Pro Tyr Tyr Tyr Asp ProAsn AspVal Asn LysVal Lys 1 1 5 5 10 10 15 15

Gly Gly

<210> <210> 46 46 <211> <211> 7 7 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9E4 CDR3 Heavy 9E4 CDR3 HeavyChain Chain

<400> <400> 46 46

Gly Gly Gly Gly Ala AlaGly GlyIle Ile AspAsp TyrTyr 1 1 5 5

<210> <210> 47 47 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9E4 CDR1 9E4 CDR1 Light LightChain Chain

<400> <400> 47 47

Lys Ser Lys Ser Ile IleGln GlnThr Thr LeuLeu LeuLeu Tyr Tyr Ser Ser Ser Gln Ser Asn Asn Lys GlnAsn LysTyr Asn LeuTyr Leu 1 1 5 5 10 10 15 15

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Ala Ala

<210> <210> 48 48 <211> <211> 7 7 <212> <212> PRT PRT <213> <213> artificial artificial <220> <220> <223> <223> 9E4 CDR2 Light 9E4 CDR2 LightChain Chain

<400> <400> 48 48

Trp Ala Trp Ala Ser Ser Ile Ile Arg Arg Lys Lys Ser Ser 1 1 5 5

<210> <210> 49 49 <211> <211> 9 9 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9E4 CDR3 9E4 CDR3 Light Light Chain Chain

<400> <400> 49 49

Gln Gln Gln Gln Tyr Tyr Tyr Tyr Ser Ser Tyr Tyr Pro Pro Leu Leu Thr Thr 1 1 5 5

<210> <210> 50 50 <211> <211> 13 13 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> GM63 Epitope GM63 Epitope126-138 126-138

<400> <400> 50 50

Glu Met Glu Met Pro Pro Ser Ser Glu Glu Glu Glu Gly Gly Tyr Tyr Gln Gln Asp Asp Tyr Tyr Glu Glu Pro Pro 1 1 5 5 10 10

<210> <210> 51 51 <211> <211> 13 13 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> GM63 CDR1 GM63 CDR1 Heavy Heavy Chain Chain

<400> <400> 51 51

Lys Ala Lys Ala Ser Ser Gly Gly Tyr Tyr Thr Thr Phe Phe Thr Thr Asn Asn Tyr Tyr Gly Gly Ile Ile Ile Ile 1 1 5 5 10 10

<210> <210> 52 52 <211> <211> 10 10 <212> <212> PRT PRT <213> <213> artificial artificial

Page28 Page 28of of 64 64

<220> <220> <223> <223> GM63 CDR2 GM63 CDR2 Heavy Heavy Chain Chain

<400> <400> 52 52

Trp Ile Trp Ile Ser Ser Ala Ala Tyr Tyr Asn Asn Gly Gly Lys Lys Thr Thr Asn Asn 1 1 5 5 10 10

<210> <210> 53 53 <211> <211> 10 10 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> GM63 CDR3 GM63 CDR3 Heavy Heavy Chain Chain

<400> <400> 53 53

Thr Arg Thr Arg Ala Ala His His Trp Trp Gly Gly Arg Arg Phe Phe Asp Asp Tyr Tyr 1 1 5 5 10 10

<210> <210> 54 54 <211> <211> 11 11 <212> <212> PRT PRT <213> <213> artificial artificial <220> <220> <223> <223> GM63 CDR1 GM63 CDR1 Light Light Chain Chain

<400> <400> 54 54

Arg Ala Arg Ala Ser Ser Gln Gln Gly Gly Ile Ile Ser Ser Ser Ser Ala Ala Leu Leu Ala Ala 1 1 5 5 10 10

<210> <210> 55 55 <211> <211> 8 8 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> GM63 CDR2 GM63 CDR2 Light Light Chain Chain

<400> <400> 55 55

Tyr Asp Tyr Asp Ala Ala Ser Ser Ser Ser Leu Leu Glu Glu Ser Ser 1 1 5 5

<210> <210> 56 56 <211> <211> 9 9 <212> <212> PRT PRT <213> <213> artificial artificial <220> <220> <223> <223> GM63 CDR3 GM63 CDR3 Light Light Chain Chain

<400> <400> 56 56

Gln Gln Gln Gln Phe Phe Lys Lys Ser Ser Tyr Tyr Pro Pro Arg Arg Thr Thr 1 1 5 5

Page29 Page 29of of 64 64

<210> <210> 57 57 <211> <211> 114 114 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> GM63 VH GM63 VH

<400> 400 57 57

Gln Val Gln Val Gln GlnLeu LeuVal Val GlnGln SerSer Gly Gly Ala Ala Glu Lys Glu Val Val Lys LysPro LysGly Pro AlaGly Ala 1 1 5 5 10 10 15 15

Ser Val Ile Ser Val IleVal ValSer Ser CysCys LysLys Ala Ala Ser Ser Gly Gly Tyr Phe Tyr Thr ThrThr PheAsn ThrTyrAsn Tyr 20 20 25 25 30 30

Gly Ile Gly Ile Ile IleTrp TrpVal Val ArgArg GlnGln Ala Ala Pro Pro Gly Gly Gly Gln Gln Leu GlyGlu LeuTrp Glu MetTrp Met 35 35 40 40 45 45

Gly Trp Gly Trp Ile IleSer SerAla Ala TyrTyr AsnAsn Gly Gly Lys Lys Thr Tyr Thr Asn Asn Ala TyrGln AlaAsn Gln LeuAsn Leu 50 50 55 55 60 60

Gln Gly Gln Gly Arg ArgVal ValThr Thr MetMet ThrThr Thr Thr Asp Asp Thr Thr Thr Ser Ser Ser ThrThr SerAla Thr TyrAla Tyr 65 65 70 70 75 75 80 80

Met Glu Met Glu Leu LeuArg ArgSer Ser LeuLeu ArgArg Ser Ser Asp Asp Asp Ala Asp Thr Thr Met AlaTyr MetTyr Tyr CysTyr Cys 85 85 90 90 95 95

Thr Arg Thr Arg Ala Ala His His Trp Trp Gly Gly Arg Arg Phe Phe Asp Asp Tyr Tyr Trp Trp Gly Gly Gln Gln Gly Gly Thr Thr Leu Leu 100 100 105 105 110 110

Val Thr Val Thr

<210> <210> 58 58 <211> <211> 107 107 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> GM63 VL GM63 VL

<400> <400> 58 58

Ala Ile Ala Ile Gln Gln Leu Leu Thr Thr Gln Gln Ser Ser Pro Pro Ser Ser Ser Ser Leu Leu Ser Ser Ala Ala Ser Ser Val Val Gly Gly 1 1 5 5 10 10 15 15

Asp Arg Asp Arg Val ValThr ThrIle Ile ThrThr CysCys Arg Arg Ala Ala Ser Gly Ser Gln Gln Ile GlySer IleSer SerAlaSer Ala 20 20 25 25 30 30

Leu Ala Leu Ala Trp TrpTyr TyrGln Gln GlnGln LysLys Pro Pro Gly Gly Lys Pro Lys Ala Ala Lys ProLeu LysLeu Leu IleLeu Ile 35 35 40 40 45 45

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Tyr Asp Tyr Asp Ala Ala Ser Ser Ser Ser Leu Leu Glu Glu Ser Ser Gly Gly Val Val Pro Pro Ser Ser Arg Arg Phe Phe Ser Ser Gly Gly 50 50 55 55 60 60

Ser Gly Ser Ser Gly SerGly GlyThr Thr AspAsp PhePhe Thr Thr Leu Leu Thr Thr Ile Ser Ile Ser SerLeu SerGln Leu ProGln Pro 65 65 70 70 75 75 80 80

Glu Asp Glu Asp Phe PheAla AlaThr ThrTyrTyr TyrTyr Cys Cys Gln Gln Gln Lys Gln Phe Phe Ser LysTyr SerPro Tyr ArgPro Arg 85 85 90 90 95 95

Thr Leu Thr Leu Gly GlyGln GlnGly Gly ThrThr LysLys Val Val Glu Glu Ile Lys Ile Lys 100 100 105 105

<210> <210> 59 59 <211> <211> 701 701 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> GM63 Heavy <223> GM63 Heavy CHain CHain Constant Constant Domain Domain

<400> <400> 59 59

Gly Cys Gly Cys Cys CysThr ThrCys Cys CysCys AlaAla Cys Cys Cys Cys Ala Gly Ala Ala Ala Gly GlyGly GlyCys Gly CysCys Cys 1 1 5 5 10 10 15 15

Cys Ala Cys Ala Thr ThrCys CysGly Gly GlyGly ThrThr Cys Cys Thr Thr Thr Cys Thr Cys Cys Cys CysAla CysCys AlaThrCys Thr 20 20 25 25 30 30

Gly Gly Gly Gly Cys CysGly GlyCys Cys CysCys CysCys Thr Thr Cys Cys Cys Cys Cys Thr Thr Cys CysAla CysAla Ala GlyAla Gly 35 35 40 40 45 45

Ala Gly Ala Gly Cys CysAla AlaCys Cys CysCys AlaAla Gly Gly Cys Cys Gly Cys Gly Gly Gly Gly CysGly GlyCys Gly AlaCys Ala 50 50 55 55 60 60

Cys Ala Cys Ala Gly GlyCys CysCys Cys GlyGly CysCys Cys Cys Cys Cys Thr Gly Thr Gly Gly Gly GlyCys GlyThr Cys GlyThr Gly 65 65 70 70 75 75 80 80

Cys Cys Cys Cys Thr ThrGly GlyGly GlyThrThr GlyGly Ala Ala Ala Ala Gly Ala Gly Gly Gly Cys AlaThr CysAla Thr CysAla Cys 85 85 90 90 95 95

Thr Thr Thr Thr Cys CysCys CysCys Cys CysCys GlyGly Ala Ala Gly Gly Cys Ala Cys Cys Cys Gly AlaThr GlyGly Thr AlaGly Ala 100 100 105 105 110 110

Cys Cys Gly Cys Cys GlyThr ThrGly Gly ThrThr CysCys Cys Cys Thr Thr Gly Gly Gly Ala Gly Ala AlaCys AlaThr Cys CysThr Cys 115 115 120 120 125 125

Thr Gly Thr Gly Gly GlyCys CysGly Gly CysCys CysCys Cys Cys Thr Thr Gly Cys Gly Ala Ala Cys CysThr CysCys Thr CysCys Cys 130 130 135 135 140 140

Page31 Page 31of of 64 64

Gly Gly Gly Gly Cys CysGly GlyThr Thr GlyGly CysCys Ala Ala Cys Cys Ala Cys Ala Cys Cys Thr CysThr ThrCys Thr CysCys Cys 145 145 150 150 155 155 160 160

Cys Cys Cys Cys Gly GlyCys CysCys Cys GlyGly ThrThr Gly Gly Cys Cys Thr Cys Thr Gly Gly Ala CysGly AlaAla Gly GlyAla Gly 165 165 170 170 175 175

Cys Ala Cys Ala Gly GlyCys CysGly Gly GlyGly CysCys Cys Cys Thr Thr Gly Ala Gly Thr Thr Cys AlaAla CysGly Ala CysGly Cys 180 180 185 185 190 190

Cys Thr Cys Thr Gly GlyAla AlaGly Gly CysCys AlaAla Gly Gly Cys Cys Gly Gly Gly Thr Thr Gly GlyThr GlyGly Thr AlaGly Ala 195 195 200 200 205 205

Cys Cys Cys Cys Gly GlyThr ThrGly Gly CysCys CysCys Cys Cys Ala Ala Gly Ala Gly Cys Cys Gly AlaCys GlyAla Cys GlyAla Gly 210 210 215 215 220 220

Cys Cys Cys Cys Thr ThrGly GlyGly Gly GlyGly CysCys Ala Ala Cys Cys Cys Ala Cys Cys Cys Gly AlaAla GlyCys Ala CysCys Cys 225 225 230 230 235 235 240 240

Thr Ala Thr Ala Cys CysAla AlaThr Thr CysCys ThrThr Gly Gly Cys Cys Ala Cys Ala Ala Ala Gly CysThr GlyGly Thr AlaGly Ala 245 245 250 250 255 255

Ala Cys Ala Cys Cys CysAla AlaCys Cys AlaAla AlaAla Gly Gly Cys Cys Cys Ala Cys Cys Cys Gly AlaCys GlyAla Cys AlaAla Ala 260 260 265 265 270 270

Cys Ala Cys Ala Cys CysCys CysAla Ala AlaAla GlyGly Gly Gly Thr Thr Gly Ala Gly Gly Gly Cys AlaAla CysAla Ala GlyAla Gly 275 275 280 280 285 285

Ala Gly Ala Gly Ala AlaGly GlyThr Thr GlyGly GlyGly Ala Ala Gly Gly Cys Cys Cys Cys Cys Ala CysAla AlaGly Ala AlaGly Ala 290 290 295 295 300 300

Gly Cys Gly Cys Thr ThrGly GlyCys Cys GlyGly AlaAla Cys Cys Ala Ala Ala Ala Ala Gly Gly Cys AlaCys CysCys Cys AlaCys Ala 305 305 310 310 315 315 320 320

Cys Ala Cys Ala Cys CysCys CysThr Thr GlyGly CysCys Cys Cys Cys Cys Cys Cys Cys Cys Cys Cys CysThr CysGly Thr CysGly Cys 325 325 330 330 335 335

Cys Cys Cys Cys Ala AlaGly GlyCys Cys CysCys CysCys Cys Cys Ala Ala Gly Gly Gly Ala Ala Cys GlyThr CysGly Thr CysGly Cys 340 340 345 345 350 350

Thr Gly Thr Gly Gly GlyGly GlyCys Cys GlyGly GlyGly Ala Ala Cys Cys Cys Ala Cys Cys Cys Gly AlaCys GlyGly Cys ThrGly Thr 355 355 360 360 365 365

Gly Thr Gly Thr Thr ThrCys CysCys Cys ThrThr GlyGly Thr Thr Thr Thr Cys Cys Cys Cys Cys Cys CysCys CysCys Cys CysCys Cys 370 370 375 375 380 380

Ala Ala Ala Ala Gly GlyCys CysCys Cys CysCys AlaAla Ala Ala Gly Gly Gly Cys Gly Ala Ala Ala CysCys AlaCys Cys CysCys Cys 385 385 390 390 395 395 400 400

Page32 Page 32of of 64 64

Thr Gly Thr Gly Ala AlaThr ThrGly Gly AlaAla ThrThr Cys Cys Ala Ala Gly Ala Gly Cys Cys Gly AlaGly GlyAla Gly CysAla Cys 405 405 410 410 415 415

Cys Cys Cys Cys Cys CysCys CysGly Gly AlaAla GlyGly Gly Gly Thr Thr Gly Cys Gly Ala Ala Cys CysThr CysGly Thr CysGly Cys 420 420 425 425 430 430

Gly Thr Gly Thr Gly GlyGly GlyThr Thr GlyGly GlyGly Thr Thr Gly Gly Gly Cys Gly Ala Ala Gly CysThr GlyGly Thr AlaGly Ala 435 435 440 440 445 445

Gly Cys Gly Cys Cys CysAla AlaCys Cys GlyGly AlaAla Gly Gly Gly Gly Ala Cys Ala Cys Cys Cys CysAla CysGly Ala AlaGly Ala 450 450 455 455 460 460

Gly Gly Gly Gly Thr ThrGly GlyAla Ala AlaAla GlyGly Thr Thr Thr Thr Cys Ala Cys Ala Ala Cys AlaThr CysGly Thr GlyGly Gly 465 465 470 470 475 475 480 480

Thr Ala Thr Ala Cys CysGly GlyThr Thr GlyGly GlyGly Ala Ala Cys Cys Gly Cys Gly Gly Gly Gly CysThr GlyGly Thr GlyGly Gly 485 485 490 490 495 495

Ala Gly Ala Gly Gly GlyThr ThrGly Gly CysCys AlaAla Cys Cys Ala Ala Ala Gly Ala Cys Cys Cys GlyCys CysAla Cys AlaAla Ala 500 500 505 505 510 510

Gly Ala Gly Ala Cys CysCys CysAla Ala AlaAla GlyGly Cys Cys Cys Cys Cys Gly Cys Ala Ala Ala GlyGly AlaAla Gly GlyAla Gly 515 515 520 520 525 525

Gly Ala Gly Ala Gly GlyCys CysAla Ala GlyGly ThrThr Ala Ala Cys Cys Ala Cys Ala Ala Ala Ala CysGly AlaCys Gly AlaCys Ala 530 530 535 535 540 540

Cys Cys Cys Cys Thr ThrAla AlaCys Cys AlaAla GlyGly Gly Gly Gly Gly Thr Gly Thr Gly Gly Thr GlyGly ThrThr Gly CysThr Cys 545 545 550 550 555 555 560 560

Cys Gly Cys Gly Thr ThrGly GlyCys Cys ThrThr GlyGly Ala Ala Cys Cys Cys Thr Cys Gly Gly Gly ThrCys GlyThr Cys GlyThr Gly 565 565 570 570 575 575

Cys Ala Cys Ala Cys CysCys CysAla Ala GlyGly GlyGly Ala Ala Cys Cys Thr Gly Thr Gly Gly Cys GlyThr CysGly Thr AlaGly Ala 580 580 585 585 590 590

Ala Cys Ala Cys Gly GlyGly GlyCys Cys AlaAla AlaAla Gly Gly Gly Gly Ala Thr Ala Ala Ala Ala ThrCys AlaAla Cys AlaAla Ala 595 595 600 600 605 605

Gly Thr Gly Thr Gly GlyCys CysAla Ala AlaAla GlyGly Gly Gly Thr Thr Cys Cys Cys Thr Thr Cys CysAla CysAla Ala CysAla Cys 610 610 615 615 620 620

Ala Ala Ala Ala Gly GlyGly GlyCys Cys CysCys CysCys Thr Thr Gly Gly Cys Ala Cys Cys Cys Gly AlaCys GlyCys Cys CysCys Cys 625 625 630 630 635 635 640 640

Cys Cys Cys Cys Ala AlaThr ThrCys Cys GlyGly AlaAla Ala Ala Ala Ala Ala Ala Ala Gly Gly Cys AlaCys CysAla Cys ThrAla Thr 645 645 650 650 655 655

Page 33 Page 33of of 64 64

Cys Ala Gly Cys Ala GlyCys CysAla Ala AlaAla GlyGly Gly Gly Cys Cys Cys Cys Ala Gly Ala Ala AlaGly GlyGly Gly CysGly Cys 660 660 665 665 670 670

Cys Ala Cys Ala Gly GlyCys CysCys Cys AlaAla CysCys Gly Gly Gly Gly Gly Gly Gly Ala Ala Cys GlyCys CysCys Cys CysCys Cys 675 675 680 680 685 685

Ala Gly Ala Gly Gly GlyThr ThrGly Gly ThrThr AlaAla Cys Cys Ala Ala Cys Cys Cys Cys Cys Thr Cys Thr 690 690 695 695 700 700

<210> <210> 60 60 <211> <211> 323 323 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> GM63 Kappa <223> GM63 Kappa Light Light Chain Chain Constant Constant Domain Domain

<400> <400> 60 60

Gly Ala Gly Ala Ala AlaCys CysThr Thr GlyGly ThrThr Gly Gly Gly Gly Cys Gly Cys Thr Thr Cys GlyAla CysCys Ala CysCys Cys 1 1 5 5 10 10 15 15

Ala Thr Ala Thr Cys CysThr ThrGly Gly ThrThr CysCys Thr Thr Thr Thr Cys Thr Cys Ala Ala Cys ThrThr CysThr Thr CysThr Cys 20 20 25 25 30 30

Cys Cys Cys Cys Gly GlyCys CysCys Cys AlaAla ThrThr Cys Cys Thr Thr Gly Thr Gly Ala Ala Gly ThrAla GlyGly Ala CysGly Cys 35 35 40 40 45 45

Ala Gly Ala Gly Thr ThrThr ThrGly Gly AlaAla AlaAla Ala Ala Thr Thr Cys Gly Cys Thr Thr Gly GlyAla GlyAla Ala CysAla Cys 50 50 55 55 60 60

Thr Gly Thr Gly Cys CysCys CysThr Thr CysCys AlaAla Gly Gly Thr Thr Gly Thr Gly Gly Gly Gly ThrThr GlyGly Thr CysGly Cys 65 65 70 70 75 75 80 80

Cys Thr Cys Thr Gly GlyCys CysThr ThrGlyGly AlaAla Ala Ala Cys Cys Ala Cys Ala Ala Ala Thr CysThr ThrCys Thr ThrCys Thr 85 85 90 90 95 95

Ala Cys Ala Cys Cys CysCys CysCys Cys CysCys GlyGly Gly Gly Gly Gly Ala Gly Ala Gly Gly Cys GlyCys CysAla Cys AlaAla Ala 100 100 105 105 110 110

Gly Gly Gly Gly Thr Thr Gly Gly Cys Cys Ala Ala Gly Gly Thr Thr Gly Gly Gly Gly Ala Ala Ala Ala Gly Gly Gly Gly Thr Thr Gly Gly 115 115 120 120 125 125

Gly Ala Gly Ala Cys CysAla AlaAla Ala CysCys GlyGly Cys Cys Cys Cys Cys Gly Cys Thr Thr Cys GlyAla CysGly Ala AlaGly Ala 130 130 135 135 140 140

Gly Cys Gly Cys Gly GlyGly GlyCys Cys AlaAla AlaAla Cys Cys Ala Ala Gly Cys Gly Cys Cys Ala CysGly AlaGly Gly AlaGly Ala 145 145 150 150 155 155 160 160

Gly Ala Gly Ala Gly Gly Cys Cys Gly Gly Thr Thr Cys Cys Ala Ala Cys Cys Cys Cys Gly Gly Ala Ala Gly Gly Cys Cys Ala Ala Gly Gly 165 165 170 170 175 175

https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H... 14/06/2019 https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H. 14/06/2019

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Gly Ala Gly Ala Cys Cys Ala Ala Gly Gly Cys Cys Ala Ala Ala Ala Gly Gly Gly Gly Ala Ala Cys Cys Thr Thr Cys Cys Cys Cys Ala Ala 180 180 185 185 190 190

Cys Cys Cys Cys Thr ThrAla AlaCys Cys AlaAla GlyGly Cys Cys Cys Cys Thr Ala Thr Gly Gly Gly AlaCys GlyAla Cys GlyAla Gly 195 195 200 200 205 205

Cys Ala Cys Ala Cys CysCys CysCys Cys ThrThr GlyGly Ala Ala Cys Cys Cys Thr Cys Cys Cys Gly ThrThr GlyCys Thr CysCys Cys 210 210 215 215 220 220

Ala Ala Ala Ala Gly GlyGly GlyCys Cys CysCys GlyGly Ala Ala Cys Cys Thr Cys Thr Ala Ala Gly CysAla GlyGly Ala AlaGly Ala 225 225 230 230 235 235 240 240

Ala Gly Ala Gly Cys CysAla AlaCys Cys AlaAla AlaAla Gly Gly Gly Gly Thr Thr Thr Gly Gly Ala ThrCys AlaGly Cys CysGly Cys 245 245 250 250 255 255

Cys Thr Cys Thr Gly GlyCys CysGly Gly AlaAla GlyGly Gly Gly Thr Thr Gly Cys Gly Ala Ala Cys CysCys CysAla Cys CysAla Cys 260 260 265 265 270 270

Cys Ala Cys Ala Gly GlyGly GlyGly Gly CysCys CysCys Thr Thr Gly Gly Thr Cys Thr Cys Cys Ala CysGly AlaCys Gly CysCys Cys 275 275 280 280 285 285

Cys Cys Cys Cys Gly GlyThr ThrGly Gly AlaAla CysCys Cys Cys Ala Ala Ala Ala Ala Gly Gly Gly AlaCys GlyThr Cys ThrThr Thr 290 290 295 295 300 300

Cys Ala Cys Ala Ala AlaCys CysAla Ala GlyGly GlyGly Gly Gly Gly Gly Cys Ala Cys Gly Gly Gly AlaThr GlyGly Thr CysGly Cys 305 305 310 310 315 315 320 320

Thr Gly Thr Gly Ala Ala

<210> <210> 61 61 <211> <211> 15 15 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9E4 Epitope126-140 9E4 Epitope 126-140

<400> <400> 61 61

Glu Met Glu Met Pro ProSer SerGlu Glu GluGlu GlyGly Tyr Tyr Gln Gln Asp Glu Asp Tyr Tyr Pro GluGlu ProAla Glu Ala 1 1 5 5 10 10 15 15

<210> <210> 62 62 <211> <211> 5 5 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR1 VH CDR1 VH

Page 35 Page 35of of 64 64

<400> <400> 62 62

Ser Tyr Ser Tyr Trp TrpMet MetHis His 1 1 5 5

<210> <210> 63 63 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR2 VH CDR2 VH

<400> <400> 63 63

Arg Ile Arg Ile Asp Asp Pro Pro Asn Asn Ser Ser Gly Gly Thr Thr Thr Thr Lys Lys Tyr Tyr Asn Asn Val Val Asn Asn Phe Phe Lys Lys 1 1 5 5 10 10 15 15

Thr Thr

<210> <210> 64 64 <211> <211> 12 12 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR3 VH CDR3 VH

<400> <400> 64 64

Leu Gly Leu Gly His His Tyr Tyr Gly Gly Asn Asn Leu Leu Tyr Tyr Ala Ala Met Met Asp Asp Tyr Tyr 1 1 5 5 10 10

<210> <210> 65 65 <211> <211> 10 10 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR1 VL CDR1 VL

<400> <400> 65 65

Ser Ala Ser Ala Ser SerSer SerSer Ser ValVal SerSer Tyr Tyr Met Met His His 1 1 5 5 10 10

<210> <210> 66 66 <211> <211> 7 7 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR2 VL CDR2 VL

<400> <400> 66 66

Asp Thr Asp Thr Ser Ser Lys Lys Leu Leu Ala Ala Ser Ser 1 1 5 5

Page36 Page 36of of 64 64

<210> <210> 67 67 <211> <211> 9 9 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR3 VL CDR3 VL

<400> <400> 67 67

Gln Gln Gln Gln Trp TrpSer SerSer Ser AsnAsn ProPro Pro Pro Thr Thr 1 1 5 5

<210> <210> 68 68 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> m2E6 VH m2E6 VH

<400> <400> 68 68

Gln Val Gln Val Gln GlnLeu LeuGln Gln GlnGln ProPro Gly Gly Ala Ala Glu Val Glu Leu Leu Lys ValPro LysGly Pro AlaGly Ala 1 1 5 5 10 10 15 15

Ser Val Lys Ser Val LysLeu LeuSer Ser CysCys ThrThr Ala Ala Ser Ser Gly Gly Tyr Phe Tyr Thr ThrThr PheSer ThrTyrSer Tyr 20 20 25 25 30 30

Trp Met Trp Met His HisTrp TrpMet Met LysLys GlnGln Arg Arg Pro Pro Gly Gly Gly Arg Arg Leu GlyGlu LeuTrp Glu IleTrp Ile 35 35 40 40 45 45

Gly Arg Gly Arg Ile IleAsp AspPro Pro AsnAsn SerSer Gly Gly Thr Thr Thr Tyr Thr Lys Lys Asn TyrVal AsnAsn Val PheAsn Phe 50 50 55 55 60 60

Lys Thr Lys Thr Lys LysAla AlaThr Thr LeuLeu ThrThr Val Val Asp Asp Lys Ser Lys Pro Pro Ser SerThr SerAla Thr TyrAla Tyr 65 65 70 70 75 75 80 80

Met His Met His Leu LeuSer SerSer Ser LeuLeu ThrThr Ser Ser Glu Glu Asp Ala Asp Ser Ser Val AlaTyr ValTyr Tyr CysTyr Cys 85 85 90 90 95 95

Ala Arg Ala Arg Leu LeuGly GlyHis His TyrTyr GlyGly Asn Asn Leu Leu Tyr Met Tyr Ala Ala Asp MetTyr AspTrp Tyr GlyTrp Gly 100 100 105 105 110 110

Gln Gly Gln Gly Thr ThrSer SerVal Val ThrThr 115 115

<210> <210> 69 69 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220>

Page37 Page 37of of 64 64

<223> <223> m2E6 VL m2E6 VL

<400> 400 69 69

Gln Ile Gln Ile Val ValLeu LeuThr Thr GlnGln SerSer Pro Pro Ala Ala Ile Ser Ile Met Met Ala SerSer AlaPro Ser GlyPro Gly 1 1 5 5 10 10 15 15

Glu Lys Glu Lys Val ValThr ThrMet Met ThrThr CysCys Ser Ser Ala Ala Ser Ser Ser Ser Ser Val SerSer ValTyr Ser MetTyr Met 20 20 25 25 30 30

His Trp His Trp Tyr TyrGln GlnGln Gln LysLys SerSer Gly Gly Thr Thr Ser Lys Ser Pro Pro Arg LysTrp ArgIle Trp TyrIle Tyr 35 35 40 40 45 45

Asp Thr Asp Thr Ser SerLys LysLeu Leu AlaAla SerSer Gly Gly Val Val Pro Arg Pro Thr Thr Phe ArgSer PheGly Ser SerGly Ser 50 50 55 55 60 60

Gly Ser Gly Ser Gly GlyThr ThrSer Ser TyrTyr SerSer Leu Leu Thr Thr Ile Ser Ile Ser Ser Met SerAsp MetThr Asp GluThr Glu 65 65 70 70 75 75 80 80

Asp Ala Asp Ala Ala AlaThr ThrTyr TyrTyrTyr CysCys Gln Gln Gln Gln Trp Ser Trp Ser Ser Asn SerPro AsnPro Pro ThrPro Thr 85 85 90 90 95 95

Phe Gly Phe Gly Ala AlaGly GlyThr Thr LysLys LeuLeu Glu Glu Leu Leu Lys Lys 100 100 105 105

<210> <210> 70 70 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> ch2E6 VH ch2E6 VH

<400> <400> 70 70

Met His Met His Leu LeuSer SerSer SerLeuLeu ThrThr Ser Ser Glu Glu Asp Ala Asp Ser Ser Val AlaTyr ValTyr Tyr CysTyr Cys 85 85 90 90 95 95

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Ala Arg Ala Arg Leu Leu Gly Gly His His Tyr Tyr Gly Gly Asn Asn Leu Leu Tyr Tyr Ala Ala Met Met Asp Asp Tyr Tyr Trp Trp Gly Gly 100 100 105 105 110 110

Gln Gly Gln Gly Thr ThrSer SerVal Val ThrThr 115 115

<210> <210> 71 71 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> ch2E6 VL ch2E6 VL

<400> <400> 71 71

Gln Ile Gln Ile Val Val Leu Leu Thr Thr Gln Gln Ser Ser Pro Pro Ala Ala Ile Ile Met Met Ser Ser Ala Ala Ser Ser Pro Pro Gly Gly 1 1 5 5 10 10 15 15

Glu Lys Glu Lys Val ValThr ThrMet Met ThrThr CysCys Ser Ser Ala Ala Ser Ser Ser Ser Ser Val SerSer ValTyr SerMetTyr Met 20 20 25 25 30 30

Gly Ser Gly Ser Gly Gly Thr Thr Ser Ser Tyr Tyr Ser Ser Leu Leu Thr Thr Ile Ile Ser Ser Ser Ser Met Met Asp Asp Thr Thr Glu Glu 65 65 70 70 75 75 80 80

<210> <210> 72 72 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 2E6-HLD1 VH 2E6-HLD1 VH

<400> <400> 72 72

Ser Val Lys Ser Val LysVal ValSer Ser CysCys LysLys Ala Ala Ser Ser Gly Gly Tyr Phe Tyr Thr ThrThr PheSer ThrTyrSer Tyr 20 20 25 25 30 30

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Trp Met Trp Met His HisTrp TrpMet Met ArgArg GlnGln Ala Ala Pro Pro Gly Gly Gly Gln Gln Leu GlyGlu LeuTrp Glu IleTrp Ile 35 35 40 40 45 45

Lys Thr Lys Thr Arg ArgAla AlaThr Thr LeuLeu ThrThr Val Val Asp Asp Lys Thr Lys Ser Ser Ser ThrThr SerAla Thr TyrAla Tyr 65 65 70 70 75 75 80 80

Met Glu Met Glu Leu LeuSer SerSer SerLeuLeu ArgArg Ser Ser Glu Glu Asp Ala Asp Thr Thr Val AlaTyr ValTyr Tyr CysTyr Cys 85 85 90 90 95 95

Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

<210> <210> 73 73 <211> <211> 107 107 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 2E6-HLD1 VL 2E6-HLD1 VL

<400> <400> 73 73

Glu Ile Glu Ile Val Val Leu Leu Thr Thr Gln Gln Ser Ser Pro Pro Ala Ala Thr Thr Leu Leu Ser Ser Leu Leu Ser Ser Pro Pro Gly Gly 1 1 5 5 10 10 15 15

Glu Arg Glu Arg Ala AlaThr ThrLeu Leu SerSer CysCys Ser Ser Ala Ala Ser Ser Ser Ser Ser Val SerSer ValTyr SerMetTyr Met 20 20 25 25 30 30

His Trp His Trp Tyr Tyr Gln Gln Gln Gln Lys Lys Pro Pro Gly Gly Gln Gln Ala Ala Pro Pro Arg Arg Arg Arg Trp Trp Ile Ile Tyr Tyr 35 35 40 40 45 45

Asp Thr Asp Thr Ser SerLys LysLeu Leu AlaAla SerSer Gly Gly Val Val Pro Arg Pro Ala Ala Phe ArgSer PheGly Ser SerGly Ser 50 50 55 55 60 60

Gly Ser Gly Ser Gly GlyThr ThrAsp Asp TyrTyr ThrThr Leu Leu Thr Thr Ile Ser Ile Ser Ser Leu SerGlu LeuPro Glu GluPro Glu 65 65 70 70 75 75 80 80

Asp Phe Asp Phe Ala AlaVal ValTyr TyrTyrTyr CysCys Gln Gln Gln Gln Trp Ser Trp Ser Ser Asn SerPro AsnPro Pro ThrPro Thr 85 85 90 90 95 95

Phe Gly Phe Gly Gln GlnGly GlyThr Thr LysLys LeuLeu Glu Glu Ile Ile Lys Arg Lys Arg 100 100 105 105

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<210> <210> 74 74 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> 2E6-HLD2 VH 2E6-HLD2 VH

<400> <400> 74 74

Trp Met Trp Met His HisTrp TrpVal Val ArgArg GlnGln Ala Ala Pro Pro Gly Gly Gly Gln Gln Leu GlyGlu LeuTrp Glu IleTrp Ile 35 35 40 40 45 45

Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

<210> 220> 75 75 <211> <211> 107 107 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 2E6-HLD2 VL 2E6-HLD2 VL

<400> <400> 75 75

Glu Ile Glu Ile Val ValLeu LeuThr Thr GlnGln SerSer Pro Pro Ala Ala Thr Ser Thr Leu Leu Leu SerSer LeuPro Ser GlyPro Gly 1 1 5 5 10 10 15 15

Glu Arg Glu Arg Ala AlaThr ThrLeu Leu SerSer CysCys Ser Ser Ala Ala Ser Ser Ser Ser Ser Val SerSer ValTyr Ser MetTyr Met 20 20 25 25 30 30

His Trp His Trp Tyr TyrGln GlnGln Gln LysLys ProPro Gly Gly Gln Gln Ala Arg Ala Pro Pro Arg ArgTrp ArgIle Trp TyrIle Tyr 35 35 40 40 45 45

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Asp Phe Asp Phe Ala AlaVal ValTyr Tyr TyrTyr CysCys Gln Gln Gln Gln Trp Ser Trp Ser Ser Asn SerPro AsnPro Pro ThrPro Thr 85 85 90 90 95 95

<210> <210> 76 76 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 2E6-HLD 2E6-HLD 3 VH 3 VH <400> <400> 76 76

Ser Val Ser Val Lys LysVal ValSer Ser CysCys LysLys Ala Ala Ser Ser Gly Gly Tyr Phe Tyr Thr ThrThr PheSer Thr TyrSer Tyr 20 20 25 25 30 30

Lys Thr Lys Thr Arg ArgVal ValThr Thr LeuLeu ThrThr Val Val Asp Asp Lys Thr Lys Ser Ser Ser ThrThr SerAla Thr TyrAla Tyr 65 65 70 70 75 75 80 80

Met Glu Met Glu Leu LeuSer SerSer Ser LeuLeu ArgArg Ser Ser Glu Glu Asp Ala Asp Thr Thr Val AlaTyr ValTyr Tyr CysTyr Cys 85 85 90 90 95 95

Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

<210> <210> 77 77 <211> <211> 107 107 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220>

Page 42 Page 42of of 64 64

<223> <223> 2E6-HLD 2E6-HLD 33 VL VL

<400> <400> 77 77

Gly Ser Gly Ser Gly Gly Thr Thr Asp Asp Tyr Tyr Thr Thr Leu Leu Thr Thr Ile Ile Ser Ser Ser Ser Leu Leu Glu Glu Pro Pro Glu Glu 65 65 70 70 75 75 80 80

Asp Phe Asp Phe Ala Ala Val Val Tyr Tyr Tyr Tyr Cys Cys Gln Gln Gln Gln Trp Trp Thr Thr Ser Ser Asn Asn Pro Pro Pro Pro Asn Asn 85 85 90 90 95 95

<210> <210> 78 78 <211> <211> 16 16 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> D1.2 CDR D1.2 CDR 1 1 Light Light Chain Chain

<400> <400> 78 78

Arg Ser Arg Ser Ser SerGln GlnSer Ser LeuLeu ValVal His His Ser Ser Asn Asn Asn Gly Gly Thr AsnTyr ThrLeu Tyr HisLeu His 1 1 5 5 10 10 15 15

<210> <210> 79 79 <211> <211> 7 7 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> D1.2 CDR D1.2 CDR 22Light LightChain Chain

<400> <400> 79 79

Lys Val Lys Val Ser SerAsn AsnArg Arg PhePhe SerSer 1 1 5 5

<210> <210> 80 80 <211> <211> 7 7 <212> <212> PRT PRT <213> <213> Artificial Artificial

Page43 Page 43of of 64 64

<220> <220> <223> D1.2 CDR <223> D1.2 CDR 33 Light Light Chain Chain

<400> <400> 80 80

Ser Gln Ser Gln Ser SerThr ThrHis His ValVal ProPro 1 1 5 5

<210> <210> 81 81 <211> <211> 13 13 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> D1.2 CDR D1.2 CDR 1 1 Heavy Heavy Chaiin Chaiin

<400> <400> 81 81

Lys Ala Lys Ala Ser Ser Gly Gly Asn Asn Thr Thr Phe Phe Thr Thr Asp Asp Tyr Tyr Glu Glu Ile Ile His His 1 1 5 5 10 10

<210> <210> 82 82 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> D1.2 CDR D1.2 CDR 2 2 Heavy Heavy Chain Chain

<400> <400> 82 82

Ala Ile Ala Ile Asp Asp Pro Pro Glu Glu Thr Thr Gly Gly Asn Asn Thr Thr Ala Ala Tyr Tyr Asn Asn Gln Gln Lys Lys Phe Phe Lys Lys 1 1 5 5 10 10 15 15

Gly Gly

<210> <210> 83 83 <211> <211> 6 6 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> D1.2 CDR D1.2 CDR 3 3 Heavy Heavy Chain Chain

<400> <400> 83 83

Ser Arg Ser Arg Gly Gly Phe Phe Asp Asp Tyr Tyr 1 1 5 5

<210> <210> 84 84 <211> <211> 219 219 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> D1.2 Light D1.2 Light Chain Chain

Page44 Page 44of of 64 64

<400> <400> 84 84

Asp Val Asp Val Met MetMet MetThr Thr GlnGln ThrThr Pro Pro Leu Leu Ser Pro Ser Leu Leu Val ProSer ValLeu Ser GlyLeu Gly 1 1 5 5 10 10 15 15

Asp Gln Asp Gln Ala AlaSer SerIle Ile SerSer CysCys Arg Arg Ser Ser Ser Ser Ser Gln Gln Leu SerVal LeuHis ValSerHis Ser 20 20 25 25 30 30

Asn Gly Asn Gly Asn AsnThr ThrTyr Tyr LeuLeu HisHis Trp Trp His His Leu Lys Leu Gln Gln Pro LysGly ProGln Gly SerGln Ser 35 35 40 40 45 45

Pro Lys Pro Lys Phe PheLeu LeuIle Ile TyrTyr LysLys Val Val Ser Ser Asn Phe Asn Arg Arg Ser PheGly SerVal Gly ProVal Pro 50 50 55 55 60 60

Asp Arg Asp Arg Phe PheSer SerGly Gly SerSer GlyGly Ser Ser Gly Gly Thr Phe Thr Asp Asp Thr PheLeu ThrLys Leu IleLys Ile 65 65 70 70 75 75 80 80

Ser Arg Ser Arg Val ValGlu GluAla AlaGluGlu AspAsp Leu Leu Gly Gly Val Phe Val Tyr Tyr Cys PheSer CysGln Ser SerGln Ser 85 85 90 90 95 95

Thr His Thr His Val ValPro ProPhe Phe ThrThr PhePhe Gly Gly Ser Ser Gly Lys Gly Thr Thr Leu LysGlu LeuIle Glu LysIle Lys 100 100 105 105 110 110

Arg Ala Arg Ala Asp Asp Ala Ala Ala Ala Pro Pro Thr Thr Val Val Ser Ser Ile Ile Phe Phe Pro Pro Pro Pro Ser Ser Ser Ser Glu Glu 115 115 120 120 125 125

Gln Leu Gln Leu Thr ThrSer SerGly Gly GlyGly AlaAla Ser Ser Val Val Val Phe Val Cys Cys Leu PheAsn LeuAsn Asn PheAsn Phe 130 130 135 135 140 140

Tyr Pro Tyr Pro Lys LysAsp AspIle Ile AsnAsn ValVal Lys Lys Trp Trp Lys Asp Lys Ile Ile Gly AspSer GlyGlu Ser ArgGlu Arg 145 145 150 150 155 155 160 160

Gln Asn Gln Asn Gly GlyVal ValLeu Leu AsnAsn SerSer Trp Trp Thr Thr Asp Asp Asp Gln Gln Ser AspLys SerAsp Lys SerAsp Ser 165 165 170 170 175 175

Thr Tyr Thr Tyr Ser SerMet MetSer Ser SerSer ThrThr Leu Leu Thr Thr Leu Lys Leu Thr Thr Asp LysGlu AspTyr Glu GluTyr Glu 180 180 185 185 190 190

Arg His Arg His Asn AsnSer SerTyr Tyr ThrThr CysCys Glu Glu Ala Ala Thr Lys Thr His His Thr LysSer ThrThr Ser SerThr Ser 195 195 200 200 205 205

Pro Ile Pro Ile Val ValLys LysSer Ser PhePhe AsnAsn Arg Arg Asn Asn Glu Cys Glu Cys 210 210 215 215

<210> <210> 85 85 <211> <211> 451 451 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220>

Page45 Page 45of of 64 64

<223> <223> D1.2 HeavyChain D1.2 Heavy Chain

<400> <400> 85 85

Gln Val Gln Val Gln Gln Leu Leu Gln Gln Gln Gln Ser Ser Gly Gly Ala Ala Glu Glu Leu Leu Val Val Arg Arg Pro Pro Gly Gly Ala Ala 1 1 5 5 10 10 15 15

Ser Val Ser Val Thr ThrLeu LeuSer Ser CysCys LysLys Ala Ala Ser Ser Gly Gly Asn Phe Asn Thr ThrThr PheAsp ThrTyrAsp Tyr 20 20 25 25 30 30

Glu Ile Glu Ile His HisTrp TrpVal Val LysLys GlnGln Thr Thr Pro Pro Val Gly Val His His Leu GlyGlu LeuTrp Glu IleTrp Ile 35 35 40 40 45 45

Gly Ala Gly Ala Ile IleAsp AspPro Pro GluGlu ThrThr Gly Gly Asn Asn Thr Tyr Thr Ala Ala Asn TyrGln AsnLys Gln PheLys Phe 50 50 55 55 60 60

Lys Gly Lys Gly Lys LysAla AlaArg Arg LeuLeu ThrThr Ala Ala Asp Asp Lys Ser Lys Ser Ser Ser SerThr SerAla Thr TyrAla Tyr 65 65 70 70 75 75 80 80

Met Glu Met Glu Leu LeuArg ArgSer SerLeuLeu ThrThr Ser Ser Glu Glu Asp Ala Asp Ser Ser Val AlaTyr ValTyr Tyr CysTyr Cys 85 85 90 90 95 95

Thr Arg Thr Arg Ser SerArg ArgGly Gly PhePhe AspAsp Tyr Tyr Trp Trp Gly Gly Gly Gln Gln Thr GlyThr ThrLeu Thr ThrLeu Thr 100 100 105 105 110 110

Val Ser Val Ser Ser Ser Ala Ala Lys Lys Thr Thr Thr Thr Pro Pro Pro Pro Ser Ser Val Val Tyr Tyr Pro Pro Leu Leu Ala Ala Pro Pro 115 115 120 120 125 125

Gly Cys Gly Cys Gly GlyAsp AspThr Thr ThrThr GlyGly Ser Ser Ser Ser Val Leu Val Thr Thr Gly LeuCys GlyLeu Cys ValLeu Val 130 130 135 135 140 140

Lys Gly Lys Gly Tyr TyrPhe PhePro Pro GluGlu SerSer Val Val Thr Thr Val Trp Val Thr Thr Asn TrpSer AsnGly Ser SerGly Ser 145 145 150 150 155 155 160 160

Leu Ser Leu Ser Ser Ser Ser Ser Val Val His His Thr Thr Phe Phe Pro Pro Ala Ala Leu Leu Leu Leu Gln Gln Ser Ser Gly Gly Leu Leu 165 165 170 170 175 175

Tyr Thr Tyr Thr Met MetSer SerSer Ser SerSer ValVal Thr Thr Val Val Pro Ser Pro Ser Ser Thr SerTrp ThrPro Trp SerPro Ser 180 180 185 185 190 190

Gln Thr Gln Thr Val ValThr ThrCys Cys SerSer ValVal Ala Ala His His Pro Ser Pro Ala Ala Ser SerThr SerThr Thr ValThr Val 195 195 200 200 205 205

Asp Lys Asp Lys Lys LysLeu LeuGlu Glu ProPro SerSer Gly Gly Pro Pro Ile Thr Ile Ser Ser Ile ThrAsn IlePro Asn CysPro Cys 210 210 215 215 220 220

Pro Pro Pro Pro Cys CysLys LysGlu Glu CysCys HisHis Lys Lys Cys Cys Pro Pro Pro Ala Ala Asn ProLeu AsnGlu Leu GlyGlu Gly 225 225 230 230 235 235 240 240

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Gly Pro Gly Pro Ser SerVal ValPhe Phe IleIle PhePhe Pro Pro Pro Pro Asn Lys Asn Ile Ile Asp LysVal AspLeu Val MetLeu Met 245 245 250 250 255 255

Ile Ser Leu Ile Ser LeuThr ThrPro Pro LysLys ValVal Thr Thr Cys Cys Val Val Val Asp Val Val ValVal AspSer Val GluSer Glu 260 260 265 265 270 270

Asp Asp Asp Asp Pro ProAsp AspVal Val ArgArg IleIle Ser Ser Trp Trp Phe Asn Phe Val Val Asn AsnVal AsnGlu Val ValGlu Val 275 275 280 280 285 285

His Thr His Thr Ala AlaGln GlnThr Thr GlnGln ThrThr His His Arg Arg Glu Tyr Glu Asp Asp Asn TyrSer AsnThr Ser IleThr Ile 290 290 295 295 300 300

Arg Val Arg Val Val ValSer SerAla Ala LeuLeu ProPro Ile Ile Gln Gln His Asp His Gln Gln Trp AspMet TrpSer Met GlySer Gly 305 305 310 310 315 315 320 320

Lys Glu Lys Glu Phe PheLys LysCys Cys LysLys ValVal Asn Asn Asn Asn Lys Leu Lys Asp Asp Pro LeuSer ProPro Ser IlePro Ile 325 325 330 330 335 335

Glu Arg Glu Arg Thr ThrIle IleSer Ser LysLys IleIle Lys Lys Gly Gly Leu Arg Leu Val Val Ala ArgPro AlaGln Pro ValGln Val 340 340 345 345 350 350

Tyr Ile Tyr Ile Leu LeuPro ProPro Pro ProPro AlaAla Glu Glu Gln Gln Leu Arg Leu Ser Ser Lys ArgAsp LysVal Asp SerVal Ser 355 355 360 360 365 365

Leu Thr Leu Thr Cys CysLeu LeuVal Val ValVal GlyGly Phe Phe Asn Asn Pro Asp Pro Gly Gly Ile AspSer IleVal Ser GluVal Glu 370 370 375 375 380 380

Trp Thr Trp Thr Ser SerAsn AsnGly Gly HisHis ThrThr Glu Glu Glu Glu Asn Lys Asn Tyr Tyr Asp LysThr AspAla Thr ProAla Pro 385 385 390 390 395 395 400 400

Val Leu Val Leu Asp AspSer SerAsp Asp GlyGly SerSer Tyr Tyr Phe Phe Ile Ser Ile Tyr Tyr Lys SerLeu LysAsp Leu IleAsp Ile 405 405 410 410 415 415

Lys Thr Lys Thr Ser SerLys LysTrp Trp GluGlu LysLys Thr Thr Asp Asp Ser Ser Ser Phe Phe Cys SerAsn CysVal Asn ArgVal Arg 420 420 425 425 430 430

His Glu His Glu Gly GlyLeu LeuLys Lys AsnAsn TyrTyr Tyr Tyr Leu Leu Lys Thr Lys Lys Lys Ile ThrSer IleArg Ser SerArg Ser 435 435 440 440 445 445

Pro Gly Pro Gly Lys Lys 450 450

<210> <210> 86 86 <211> <211> 11 11 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> C10.2 CDR C10.2 CDR1 1Light Light Chain Chain

Page 47 Page 47of of 64 64

<400> <400> 86 86

Gln Ala Gln Ala Ser Ser Gln Gln Gly Gly Thr Thr Ser Ser Ile Ile Asn Asn Leu Leu Asn Asn 1 1 5 5 10 10

<210> <210> 87 87 <211> <211> 7 7 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> C10.2 CDR C10.2 CDR 2 2 Light Light Chain Chain

<400> <400> 87 87

Gly Ala Gly Ala Ser Ser Asn Asn Leu Leu Glu Glu Asp Asp 1 1 5 5

<210> <210> 88 88 <211> <211> 7 7 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> C10.2 CDR C10.2 CDR 3 3 Light Light Chain Chain

<400> <400> 88 88

Leu Gln Leu Gln His His Thr Thr Tyr Tyr Leu Leu Pro Pro 1 1 5 5

<210> <210> 89 89 <211> <211> 13 13 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> <223> C10.2 CDR C10.2 CDR 1 1 Heavy Heavy Chain Chain

<400> <400> 89 89

Lys Ala Lys Ala Ser SerGly GlyTyr Tyr ThrThr PhePhe Thr Thr Asp Asp Arg Ile Arg Thr Thr His Ile His 1 1 5 5 10 10

<210> <210> 90 90 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> Artificial Artificial <220> <220> <223> C10.2 CDR <223> C10.2 CDR 22 Heavy Heavy Chain Chain

<400> <400> 90 90

Tyr Ile Tyr Ile Tyr Tyr Pro Pro Gly Gly Asp Asp Gly Gly Ser Ser Thr Thr Lys Lys Tyr Tyr Asn Asn Glu Glu Asn Asn Phe Phe Lys Lys 1 1 5 5 10 10 15 15

Gly Gly

Page48 Page 48of of 64 64

<210> <210> 91 91 <211> <211> 6 6 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> C10.2 CDR C10.2 CDR3 3Heary Heary Chain Chain

<400> <400> 91 91

Arg Gly Arg Gly Ala AlaMet MetAsp Asp TyrTyr 1 1 5 5

<210> <210> 92 92 <211> <211> 214 214 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> C10.2 Light C10.2 Light Chain Chain

<400> <400> 92 92

Asp Val Asp Val Gln Gln Met Met Ile Ile Gln Gln Ser Ser Pro Pro Ser Ser Ser Ser Leu Leu Ser Ser Ala Ala Ser Ser Leu Leu Gly Gly 1 1 5 5 10 10 15 15

Asp Ile Asp Ile Val Val Thr Thr Met Met Thr Thr Cys Cys Gln Gln Ala Ala Ser Ser Gln Gln Gly Gly Thr Thr Ser Ser Ile Ile Asn Asn 20 20 25 25 30 30

Leu Asn Leu Asn Trp TrpPhe PheGln Gln GlnGln LysLys Pro Pro Gly Gly Lys Pro Lys Ala Ala Lys ProLeu LysLeu Leu IleLeu Ile 35 35 40 40 45 45

Tyr Gly Tyr Gly Ala Ala Ser Ser Asn Asn Leu Leu Glu Glu Asp Asp Gly Gly Val Val Pro Pro Ser Ser Arg Arg Phe Phe Ser Ser Gly Gly 50 50 55 55 60 60

Ser Arg Tyr Ser Arg TyrGly GlyThr Thr AspAsp PhePhe Thr Thr Leu Leu Thr Thr Ile Ser Ile Ser SerLeu SerGlu Leu AspGlu Asp 65 65 70 70 75 75 80 80

Glu Asp Glu Asp Met MetAla AlaThr ThrTyrTyr PhePhe Cys Cys Leu Leu Gln Thr Gln His His Tyr ThrLeu TyrPro Leu PhePro Phe 85 85 90 90 95 95

Thr Phe Thr Phe Gly GlySer SerGly Gly ThrThr LysLys Leu Leu Glu Glu Ile Arg Ile Lys Lys Ala ArgAsp AlaAla Asp AlaAla Ala 100 100 105 105 110 110

Pro Thr Pro Thr Val ValSer SerIle Ile PhePhe ProPro Pro Pro Ser Ser Ser Ser Glu Leu Glu Gln GlnThr LeuSer Thr GlySer Gly 115 115 120 120 125 125

Gly Ala Gly Ala Ser SerVal ValVal Val CysCys PhePhe Leu Leu Asn Asn Asn Tyr Asn Phe Phe Pro TyrLys ProAsp Lys IleAsp Ile 130 130 135 135 140 140

Asn Val Asn Val Lys Lys Trp Trp Lys Lys Ile Ile Asp Asp Gly Gly Ser Ser Glu Glu Arg Arg Gln Gln Asn Asn Gly Gly Val Val Leu Leu 145 145 150 150 155 155 160 160

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Asn Ser Asn Ser Trp TrpThr ThrAsp Asp GlnGln AspAsp Ser Ser Lys Lys Asp Thr Asp Ser Ser Tyr ThrSer TyrMet Ser SerMet Ser 165 165 170 170 175 175

Ser Thr Ser Thr Leu LeuThr ThrLeu Leu ThrThr LysLys Asp Asp Glu Glu Tyr Arg Tyr Glu Glu His ArgAsn HisSer Asn TyrSer Tyr 180 180 185 185 190 190

Thr Cys Thr Cys Glu GluAla AlaThr Thr HisHis LysLys Thr Thr Ser Ser Thr Pro Thr Ser Ser Ile ProVal IleLys Val SerLys Ser 195 195 200 200 205 205

Phe Asn Phe Asn Arg ArgAsn AsnGlu Glu CysCys 210 210

<210> <210> 93 93 <211> <211> 439 439 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> C10.2 C10. 2 Heavy Chain Heavy Chain

<400> <400> 93 93

Gln Val Gln Val Gln GlnLeu LeuGln Gln GlnGln SerSer Asp Asp Ala Ala Glu Val Glu Leu Leu Lys ValPro LysGly Pro AlaGly Ala 1 1 5 5 10 10 15 15

Ser Val Ser Val Lys LysIle IleSer Ser CysCys LysLys Ala Ala Ser Ser Gly Thr Gly Tyr Tyr Phe ThrThr PheAsp Thr ArgAsp Arg 20 20 25 25 30 30

Thr Ile Thr Ile His HisTrp TrpVal Val LysLys GlnGln Arg Arg Pro Pro Glu Gly Glu Gln Gln Leu GlyGlu LeuTrp Glu IleTrp Ile 35 35 40 40 45 45

Gly Tyr Gly Tyr Ile IleTyr TyrPro Pro GlyGly AspAsp Gly Gly Ser Ser Thr Tyr Thr Lys Lys Asn TyrGlu AsnAsn Glu PheAsn Phe 50 50 55 55 60 60

Lys Gly Lys Gly Lys LysAla AlaThr Thr LeuLeu ThrThr Ala Ala Asp Asp Lys Ser Lys Ser Ser Ser SerThr SerAla Thr TyrAla Tyr 65 65 70 70 75 75 80 80

Met Gln Met Gln Leu LeuAsn AsnSer SerLeuLeu ThrThr Ser Ser Glu Glu Asp Ala Asp Ser Ser Val AlaTyr ValPhe Tyr CysPhe Cys 85 85 90 90 95 95

Ala Arg Ala Arg Arg ArgGly GlyAla Ala MetMet AspAsp Tyr Tyr Trp Trp Gly Gly Gly Gln Gln Thr GlySer ThrVal Ser ThrVal Thr 100 100 105 105 110 110

Val Ser Val Ser Ser SerAla AlaLys Lys ThrThr ThrThr Pro Pro Pro Pro Ser Tyr Ser Val Val Pro TyrLeu ProAla Leu ProAla Pro 115 115 120 120 125 125

Gly Ser Gly Ser Ala AlaAla AlaGln Gln ThrThr AsnAsn Ser Ser Met Met Val Leu Val Thr Thr Gly LeuCys GlyLeu Cys ValLeu Val 130 130 135 135 140 140

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Lys Gly Lys Gly Tyr TyrPhe PhePro Pro GluGlu ProPro Val Val Thr Thr Val Trp Val Thr Thr Asn TrpSer AsnGly Ser SerGly Ser 145 145 150 150 155 155 160 160

Leu Ser Leu Ser Ser Ser Gly Gly Val Val His His Thr Thr Phe Phe Pro Pro Ala Ala Val Val Leu Leu Gln Gln Ser Ser Asp Asp Leu Leu 165 165 170 170 175 175

Tyr Thr Tyr Thr Leu LeuSer SerSer Ser SerSer ValVal Thr Thr Val Val Pro Ser Pro Ser Ser Thr SerTrp ThrPro Trp SerPro Ser 180 180 185 185 190 190

Glu Thr Glu Thr Val ValThr ThrCys Cys AsnAsn ValVal Ala Ala His His Pro Ser Pro Ala Ala Ser SerThr SerLys Thr ValLys Val 195 195 200 200 205 205

Asp Lys Asp Lys Lys LysIle IleVal Val ProPro ArgArg Asp Asp Cys Cys Gly Lys Gly Cys Cys Pro LysCys ProIle Cys CysIle Cys 210 210 215 215 220 220

Thr Val Thr Val Pro ProGlu GluVal Val SerSer SerSer Val Val Phe Phe Ile Pro Ile Phe Phe Pro ProLys ProPro Lys LysPro Lys 225 225 230 230 235 235 240 240

Asp Val Asp Val Leu LeuThr ThrIle Ile ThrThr LeuLeu Thr Thr Pro Pro Lys Thr Lys Val Val Cys ThrVal CysVal Val ValVal Val 245 245 250 250 255 255

Asp Ile Asp Ile Ser SerLys LysAsp Asp AspAsp ProPro Glu Glu Val Val Gln Ser Gln Phe Phe Trp SerPhe TrpVal Phe AspVal Asp 260 260 265 265 270 270

Asp Val Asp Val Glu GluVal ValHis His ThrThr AlaAla Gln Gln Thr Thr Gln Arg Gln Pro Pro Glu ArgGlu GluGln Glu PheGln Phe 275 275 280 280 285 285

Asn Ser Asn Ser Thr Thr Phe Phe Arg Arg Ser Ser Val Val Ser Ser Glu Glu Leu Leu Pro Pro Ile Ile Met Met His His Gln Gln Asp Asp 290 290 295 295 300 300

Trp Leu Trp Leu Asn AsnGly GlyLys Lys GluGlu PhePhe Lys Lys Cys Cys Arg Asn Arg Val Val Ser AsnAla SerAla Ala PheAla Phe 305 305 310 310 315 315 320 320

Pro Ala Pro Ala Pro ProIle IleGlu Glu LysLys ThrThr Ile Ile Ser Ser Lys Lys Lys Thr Thr Gly LysArg GlyPro Arg LysPro Lys 325 325 330 330 335 335

Ala Pro Ala Pro Gln GlnVal ValTyr Tyr ThrThr IleIle Pro Pro Pro Pro Pro Glu Pro Lys Lys Gln GluMet GlnAla Met LysAla Lys 340 340 345 345 350 350

Asp Lys Asp Lys Val ValSer SerLeu Leu ThrThr CysCys Met Met Ile Ile Thr Phe Thr Asp Asp Phe PhePro PheGlu Pro AspGlu Asp 355 355 360 360 365 365

Ile Thr Val Ile Thr ValGlu GluTrp Trp Gln Gln TrpTrp Asn Asn Gly Gly Gln Gln Pro Glu Pro Ala AlaAsn GluTyr Asn Tyr Lys Lys 370 370 375 375 380 380

Asn Thr Asn Thr Gln GlnPro ProIle Ile MetMet AspAsp Thr Thr Asp Asp Gly Tyr Gly Ser Ser Phe TyrVal PheTyr Val SerTyr Ser 385 385 390 390 395 395 400 400

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Lys Leu Lys Leu Asn AsnVal ValGln Gln LysLys SerSer Asn Asn Trp Trp Glu Gly Glu Ala Ala Asn GlyThr AsnPhe Thr ThrPhe Thr 405 405 410 410 415 415

Cys Ser Cys Ser Val ValLeu LeuHis His GluGlu GlyGly Leu Leu His His Asn His Asn His His Thr HisGlu ThrLys Glu SerLys Ser 420 420 425 425 430 430

Leu Ser Leu Ser His HisSer SerPro Pro GlyGly LysLys 435 435

<210> <210> 94 94 <211> <211> 10 10 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR1 VL CDR1 VL 7C4 7C4

<400> <400> 94 94

Ser Ala Ser Ala Ser SerSer SerSer Ser ValVal SerSer Phe Phe Met Met His His 1 1 5 5 10 10

<210> <210> 95 95 <211> <211> 10 10 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR1 VL CDR1 VL 7A10/8D9 7A10/8D9

<400> <400> 95 95

Ser Ala Ser Ala Ser SerSer SerSer Ser ValVal SerSer Tyr Tyr Ile Ile His His 1 1 5 5 10 10

<210> <210> 96 96 <211> <211> 9 9 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR3 VL CDR3 VL L3 L3

<400> <400> 96 96

Gln Gln Gln Gln Trp TrpThr ThrSer Ser AsnAsn ProPro Pro Pro Phe Phe 1 1 5 5

<210> <210> 97 97 <211> <211> 5 5 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR1 VH CDR1 VH 7C4 7C4

<400> <400> 97 97

https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H... 14/06/2019 https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H.. 14/06/2019

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Arg Tyr Arg Tyr Trp TrpMet MetHis His 1 1 5 5

<210> <210> 98 98 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR2 VH CDR2 VH 5A1 5A1

<400> <400> 98 98

Arg Val Arg Val Asp AspPro ProAsn Asn SerSer GlyGly Thr Thr Thr Thr Lys Asn Lys Tyr Tyr Val AsnAsn ValPhe Asn LysPhe Lys 1 1 5 5 10 10 15 15

Thr Thr

<210> <210> 99 99 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR2 VH CDR2 VH 9G11 9G11

<400> <400> 99 99

Arg Ile Arg Ile Asp AspPro ProAsn Asn SerSer GlyGly Thr Thr Thr Thr Lys Asn Lys Tyr Tyr Val AsnHis ValPhe His LysPhe Lys 1 1 5 5 10 10 15 15

Thr Thr

<210> <210> 100 100 <211> <211> 17 17 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR2 VH CDR2 VH 9C12 9C12

<400> <400> 100 100

Arg Ile Arg Ile Asp AspPro ProAsn Asn SerSer GlyGly Thr Thr Thr Thr Lys Asn Lys Tyr Tyr Val AsnAsn ValIle Asn LysIle Lys 1 1 5 5 10 10 15 15

Thr Thr

<210> <210> 101 101 <211> <211> 12 12 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220>

Page53 Page 53of of 64 64

<223> <223> CDR3 VH CDR3 VH 5A1 5A1

<400> <400> 101 101

Leu Gly Leu Gly His HisTyr TyrGly Gly AsnAsn LeuLeu Asn Asn Ala Ala Met Tyr Met Asp Asp Tyr 1 1 5 5 10 10

<210> <210> 102 102 <211> <211> 12 12 <212> <212> PRT PRT <213> <213> Artificial Artificial

<220> <220> <223> <223> CDR3 VH CDR3 VH 9D7 9D7

<400> <400> 102 102

Leu Gly Leu Gly His HisTyr TyrSer Ser LysLys ValVal Leu Leu Ala Ala Met Tyr Met Asp Asp Tyr 1 1 5 5 10 10

<210> <210> 103 103 <211> <211> 12 12 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> CDR3 VH CDR3 VH 7A10/8D9 7A10/8D9

<400> <400> 103 103

Leu Gly Leu Gly His HisTyr TyrGly Gly AsnAsn LeuLeu Tyr Tyr Ala Ala Lys Tyr Lys Asp Asp Tyr 1 1 5 5 10 10

<210> <210> 104 104 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 5A1 VL 5A1 VL

<400> <400: 104 104

Gly Ser Gly Ser Gly Gly Thr Thr Asp Asp Tyr Tyr Thr Thr Leu Leu Thr Thr Ile Ile Ser Ser Arg Arg Leu Leu Glu Glu Pro Pro Glu Glu 65 65 70 70 75 75 80 80

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Phe Gly Phe Gly Gln GlnGly GlyThr Thr LysLys LeuLeu Glu Glu Ile Ile Lys Lys 100 100 105 105

<210> <210> 105 105 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 5A1 VH 5A1 VH

<400> <400> 105 105

Gln Val Gln Val Gln GlnLeu LeuVal Val GlnGln SerSer Gly Gly Ala Ala Glu Lys Glu Val Val Lys LysGln LysGly Gln AlaGly Ala 1 1 5 5 10 10 15 15

Ser Val Ser Val Lys LysVal ValSer Ser CysCys LysLys Ala Ala Ser Ser Gly Gly Tyr Phe Tyr Thr ThrThr PheSer ThrTyrSer Tyr 20 20 25 25 30 30

Trp Met Trp Met His HisTyr TyrMet Met ArgArg GlnGln Ala Ala Pro Pro Gly Gly Gly Gln Gln Leu GlyGlu LeuTrp Glu IleTrp Ile 35 35 40 40 45 45

Gly Arg Gly Arg Val ValAsp AspPro Pro AsnAsn SerSer Gly Gly Thr Thr Thr Tyr Thr Lys Lys Asn TyrVal AsnAsn Val PheAsn Phe 50 50 55 55 60 60

Lys Thr Lys Thr Arg ArgAla AlaThr Thr LeuLeu ThrThr Val Val Asp Asp Lys Thr Lys Thr Thr Ser ThrThr SerAla Thr TyrAla Tyr 65 65 70 70 75 75 80 80

Ala Arg Ala Arg Leu Leu Gly Gly His His Tyr Tyr Gly Gly Asn Asn Leu Leu Asn Asn Ala Ala Met Met Asp Asp Tyr Tyr Trp Trp Gly Gly 100 100 105 105 110 110

Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

<210> <210> 106 106 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9D7 VL 9D7 VL

<400> <400> 106 106

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<210> <210> 107 107 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9D7 VH 9D7 VH

<400> 400> 107 107

Ala Arg Ala Arg Leu Leu Gly Gly His His Tyr Tyr Ser Ser Lys Lys Val Val Leu Leu Ala Ala Met Met Asp Asp Tyr Tyr Trp Trp Gly Gly 100 100 105 105 110 110

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Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

<210> <210> 108 108 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> 223> 9G11 VL 9G11 VL

<400> <4001 108 108

His Trp His Trp Tyr TyrGln GlnGln Gln LysLys GlnGln Gly Gly Gln Gln Ala Arg Ala Pro Pro Arg ArgTrp ArgIle Trp TyrIle Tyr 35 35 40 40 45 45

<210> <210> 109 109 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9G11 VH 9G11 VH

<400> <400> 109 109

Page 57 Page 57of of 64 64

Gly Arg Gly Arg Ile IleAsp AspPro Pro AsnAsn SerSer Gly Gly Thr Thr Thr Tyr Thr Lys Lys Asn TyrVal AsnHis Val PheHis Phe 50 50 55 55 60 60

Ala Arg Ala Arg Leu Leu Gly Gly His His Tyr Tyr Gly Gly Asn Asn Leu Leu Tyr Tyr Ala Ala Thr Thr Asp Asp Tyr Tyr Trp Trp Gly Gly 100 100 105 105 110 110

Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

<210> <210> 110 110 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 7C4 VL 7C4 VL

<400> 400> 110 110

Glu Arg Glu Arg Ala AlaThr ThrLeu Leu SerSer CysCys Ser Ser Ala Ala Ser Ser Ser Ser Ser Val SerSer ValPhe Ser MetPhe Met 20 20 25 25 30 30

<210> <210> 111 111 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220>

Page58 Page 58of of 64 64

<223> 223> 7C4 VH 7C4 VH

<400> <400> 111 111

Ser Val Lys Ser Val LysVal ValSer Ser CysCys LysLys Ala Ala Ser Ser Gly Gly Tyr Phe Tyr Thr ThrThr PheArg ThrTyrArg Tyr 20 20 25 25 30 30

Trp Met Trp Met His HisTrp TrpMet Met ArgArg GlnGln Ala Ala Pro Pro Gly Gly Gly Gln Gln Pro GlyGlu ProTrp Glu IleTrp Ile 35 35 40 40 45 45

Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

<210> <210> 112 112 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> L3 VL L3 VL

<400> <400> 112 112

Page59 Page 59of of 64 64

<210> <210> 113 113 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> L3 VH L3 VH

<400> <400> 113 113

Gln Val Gln Val Gln GlnLeu LeuVal Val GlnGln GlnGln Gly Gly Ala Ala Glu Lys Glu Val Val Lys LysPro LysGly Pro AlaGly Ala 1 1 5 5 10 10 15 15

Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

<210> <210> 114 114 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 7A10 VL 7A10 VL

<400> <400> 114 114

Page60 Page 60of of 64 64

Glu Arg Glu Arg Ala AlaThr ThrLeu Leu SerSer CysCys Ser Ser Ala Ala Ser Ser Ser Ser Ser Val SerSer ValTyr Ser IleTyr Ile 20 20 25 25 30 30

Asp Phe Asp Phe Ala AlaVal ValTyr TyrTyrTyr CysCys Gln Gln Gln Gln Trp Ser Trp Thr Thr Asn SerPro AsnPro Pro AsnPro Asn 85 85 90 90 95 95

<210> <210> 115 115 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 7A10 VH 7A10 VH

<400> <400> 115 115

Ala Arg Ala Arg Leu Leu Gly Gly His His Tyr Tyr Gly Gly Asn Asn Leu Leu Tyr Tyr Ala Ala Lys Lys Asp Asp Tyr Tyr Trp Trp Gly Gly 100 100 105 105 110 110

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Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

<210> <210> 116 116 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 8D9 VL 8D9 VL

<400> <400: 116 116

<210> <210> 117 117 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 8D9 VH 8D9 VH

<400> <400> 117 117

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Ala Arg Ala Arg Leu LeuGly GlyHis His TyrTyr GlyGly Asn Asn Leu Leu Tyr Lys Tyr Ala Ala Asp LysTyr AspTrp Tyr GlyTrp Gly 100 100 105 105 110 110

Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

<210> <210> 118 118 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 9C12 VL 9C12 VL

<400> :400> 118 118

His Trp His Trp Tyr TyrGln GlnGln Gln LysLys ProPro Gly Gly Gln Gln Ala Arg Ala Pro Pro Arg ArgLeu ArgIle Leu TyrIle Tyr 35 35 40 40 45 45

<210> <210> 119 119 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220>

Page63 Page 63of of 64 64

<223> :223: > 9C12 VH 9C12 VH

<400> :400> 119 119

Gly Arg Gly Arg Ile IleAsp AspPro Pro AsnAsn SerSer Gly Gly Thr Thr Thr Tyr Thr Lys Lys Asn TyrVal AsnAsn Val IleAsn Ile 50 50 55 55 60 60

Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

<210> <210> 120 120 <211> <211> 106 106 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 6B6 VL 6B6 VL

<400> <400> 120 120

https://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H... 14/06/2019 ttps://patentscope.wipo.int/search/docs2/pct/WO2018115225/file/NnXhGXrLbqO7H..14/06/2019

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<210> <210> 121 121 <211> <211> 118 118 <212> <212> PRT PRT <213> <213> artificial artificial

<220> <220> <223> <223> 6B6 VH 6B6 VH

<400> <400> 121 121

Gln Gly Gln Gly Thr ThrLeu LeuVal Val ThrThr 115 115

Claims

CLAIMS 1. Use of an alpha-synuclein binding monoclonal antibody, or an antigen binding fragment thereof, in the manufacture of a medicament for inhibit ing aggregation of tau, wherein said alpha-synuclein binding antibody or antigen-binding fragment thereof is selected from the group consisting of (A)-(D):

(A) a monoclonal antibody, or antigen-binding fragment thereof, comprising: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:2; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6;

(B) a monoclonal antibody, or antigen-binding fragment thereof, comprising: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:33; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4;

(e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6;

(C) a monoclonal antibody, or antigen-binding fragment thereof, comprising: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:34; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6;

(D) a monoclonal antibody, or antigen-binding fragment thereof, comprising: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:35; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and

(f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6.

2. A method of inhibiting aggregation of tau which comprises administering to a subject an effective amount of an alpha-synuclein binding monoclonal antibody, or an antigen-binding fragment thereof, wherein said alpha synuclein binding antibody or antigen-binding fragment thereof is selected from the group consisting of (A)-(D):

(A) a monoclonal antibody, or antigen-binding fragment thereof, comprising: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:2; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6;

(B) a monoclonal antibody, or antigen-binding fragment thereof, comprising: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:33; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3;

(d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6;

(C) a monoclonal antibody, or antigen-binding fragment thereof, comprising: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:34; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6;

(D) a monoclonal antibody, or antigen-binding fragment thereof, comprising: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:35; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4;

(e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6.

3. The use of claim 1 or the method of claim 2, wherein the monoclonal antibody, or antigen-binding fragment thereof comprises: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:2; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6.

4. The use of claim 1 or the method of claim 2, wherein the monoclonal antibody, or antigen-binding fragment thereof comprises: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:33; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and

(f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6.

5. The use of claim 1 or the method of claim 2, wherein the monoclonal antibody, or antigen-binding fragment thereof comprises: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:34; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6.

6. The use of claim 1 or the method of claim 2, wherein the monoclonal antibody, or antigen-binding fragment thereof comprises: (a) a Heavy Chain CDR1 having the amino acid sequence of SEQ ID NO:1; (b) a Heavy Chain CDR2 having the amino acid sequence of SEQ ID NO:35; (c) a Heavy Chain CDR3 having the amino acid sequence of SEQ ID NO:3; (d) a Light Chain CDR1 having the amino acid sequence of SEQ ID NO:4; (e) a Light Chain CDR2 having the amino acid sequence of SEQ ID NO:5; and (f) a Light Chain CDR3 having the amino acid sequence of SEQ ID NO:6.

7. The use of any one of claims 1 or 3 to 6 or the method of any one of claims 2 to 6, wherein said alpha-synuclein antibody is for treating a tauopathy selected from the group comprising Argyrophilic Grain Disease (AGD), Psychosis, particularly Psychosis due to AD or Psychosis in pa tients with AD, psychiatric symptoms of patients with Lewy body demen tia, Progressive Supranuclear Palsy (PSP), Frontotemporal dementia (FTD or variants thereof), TBI (traumatic brain injury, acute or chronic), Corticobasal Degeneration (CBD), Picks Disease, Primary age-related tauopathy (PART), Neurofibrillary tangle-predominant senile dementia, Dementia pugilistica, Chronic traumatic encephalopathy, stroke, stroke re covery, neurodegeneration in relation to Parkinson's disease, Parkinson ism linked to chromosome, Lytico-Bodig disease (Parkinson-dementia complex of Guam), Ganglioglioma and gangliocytoma, Meningioangioma tosis, Postencephalitic parkinsonism, Subacute sclerosing panencephali tis, Huntington's disease, lead encephalopathy, tuberous sclerosis, Hal lervorden-Spatz disease and lipofuscinosis. More typically, the taupathy is selected from the group consisting of Alzheimer's disease, Argyrophilic Grain Disease (AGD), Psychosis, particularly Psychosis due to AD or Psychosis in patients with AD, psychiatric symptoms of patients with Lewy body dementia, Progressive Supranuclear Palsy (PSP), Frontotemporal dementia (FTD or variants thereof), TBI (traumatic brain injury, acute or chronic), Corticobasal Degeneration (CBD), and Picks Disease.

8. A pharmaceutical composition comprising the monoclonal antibody, or an antigen-binding fragment thereof, as recited in any one of the previous claims, and a pharmaceutically acceptable carrier.

H. Lundbeck A/S

Patent Attorneys for the Applicant/Nominated Person

SPRUSON&FERGUSON

Antibody Immunogen Protocol Mouse strain

GM37 FL human Complete HCo17-Balb/c Alpha- Freunds synuclein fibrils adjuvant and imcomplete Alternating with Freunds 1-60 alpha- adjuvant

synuclein

1-119 alpha- synuclein

GM285 FL human Complete HCo12-Balb/c alpha- Freunds synuclein adjuvant and imcomplete monomer followed by Freunds fibrils or adjuvant

monomer

FIG 1

LILL

-Graph Cell Line = 1.2x10+8 LSB6004-A-SYN-BAP 9x10+7 EC50 = 0.0006

6x10+7

3x10+7

0 -5 -3 -1

Conc

Fig 2A

Human a-syn

3.00

2.40

1.80 mm

1.20

0.60

0.00

0 400 800 1,200 1,600 2,000

Time (sec)

Human B-syn

0.60

0.40

0.20

0.00

400 800 1,200 1,600 2,000 0

Time (sec)

Fig 2B cyno a-syn

3.00

2.00

1.00

0.00

0 400 800 1,200 1,600 2,000

Time (sec)

Human y-syn 0.09

0.06

0.03 mm

0.00

0.03

0.06

400 800 1,200 1,600 2,000 0

Time (sec)

Fig 2B (continued) mouse a-syn

3.00

2.00

mm

1.00

0.00

0 400 800 1,200 1,600 2,000

Time (sec)

Fig 2B (continued)

Human a-syn A-SYN: lgG-6004-285

5.00

4 50

4 00

3 50

3 00

mu 2 50

2 00

150 100 0 50

0.00

0 50

1.000 1 200 1.400 1,600 1,800 2.000 0 200 400 600 800 Time (sec)

Fig 2C

Human B-syn B-SYN: IgG-6004-285 5.00 4.50 4.00

3.50

3.00 mu 2 50

2.00

1.50

1.00

0.50 0.00 -0 50

800 1.000 1,200 1,400 1.600 1,800 2,000 o 200 400 600 Time (sec)

Fig 2C (continued) cyno a-syn A-SYN-MF: IgG-6004-285 5.00

4.50

4.00

3.50

3.00

2.50

2.00

1 50

1.00

0.50

0.00

-0.50

200 400 600 800 1.000 1,200 1,400 1.600 1,800 2,000 o Time (sec)

Fig 2C (continued)

Human y-syn G-SYN: IgG-6004-285 5.00

4.50

4.00

3.50

3.00

2 50

2.00

1.50

1.00

0.50

0.00

-0.50

0 200 400 600 800 1.000 1.200 1,400 1,600 1,800 2.000

Time (sec)

Fig 2C (continued) mouse a-syn A-SYN-MM: IgG-6004-285 5.00

4 50

4.00

3 50

3.00

2 50

2 00

1.50

100 0.50

0.00

-0 50

0 200 400 600 800 1,000 1,200 1,400 1.600 1,800 2,000

Time (sec)

Fig 2C (continued)

WO 11/22

RW A

031

FIG

B 037 60 50 40 30 20 10 0 0 2e-8 4e-8 6e-8 8e-8 1e-7 1.2e-7

Conc

C KA(1/M) KD(nM) Rmax Chi2 Name hlgG1-6004-037 C106S 2.33E+08 4.29 61.1 0.96

Fig 3 (continued)